Histone acetyltransferase p300/CBP regulates reproductive diapause via the juvenile hormone pathway in the cabbage beetle, Colaphellus bowringi
Data files
Jul 14, 2025 version files 663.61 MB
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IMB_all_data.zip
663.57 MB
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README.md
41.42 KB
Abstract
Juvenile hormone (JH) serves as a pivotal endocrine regulator of reproductive diapause in insects, though the molecular mechanisms governing JH signaling remain poorly characterized. This investigation assesses the roles of the histone acetyltransferase p300/CBP in mediating reproductive diapause through JH signaling in the cabbage beetle Colaphellus bowringi. RNA interference-mediated suppression of p300/CBP in reproductive female adults induced diapause-like phenotypes, including arrested ovarian maturation, suppressed vitellogenesis, enhanced lipid storage, and upregulated expression of diapause-associated genes. Along with downregulation of key JH biosynthetic enzymes and signaling components. While exogenous JH application decreased lipid accumulation, it failed to rescue ovarian developmental arrest. Parallel experiments in diapausing females with p300/CBP knockdown yielded similar results - JH analog treatment reduced lipid reserves without stimulating ovarian development. These findings collectively demonstrate that p300/CBP regulates photoperiodic reproductive diapause through dual regulation of JH biosynthesis and signal transduction pathways in C. bowringi. Our results establish p300/CBP as a critical mediator of JH biosynthesis and signaling during photoperiod-induced reproductive diapause. This work advances our understanding of epigenetic regulation in insect diapause and provides a molecular framework for future studies on environmental adaptation strategies in insects.
General information
This dataset does not include large-scale datasets such as transcriptomes; instead, it is organized according to the figures and tables in the article. It primarily comprises RT-qPCR data and various biochemical assay results. The calculated data from RT-qPCR, TG, TL, WB, ovarian size measurements, lipid droplet size measurements, and HPLC/MSMS are provided in CSV format, while other incompatible data are presented in PPT files. Additionally, anatomical images of the cabbage beetle ovaries are included.
Corresponding author information
Name: Xiao-Ping Wang
ORCID: https://orcid.org/0000-0002-1718-1026
Affiliation: Hubei Key Laboratory of Insect Resources Utilization and Sustainable Pest Management, College of Plant Science and Technology, Huazhong Agricultural University, Wuhan 430070, China.
email: xpwang@mail.hzau.edu.cn
Alternative contact information
Name: Hao-Min An
ORCID: https://orcid.org/0009-0002-3615-598X
Affiliation: Hubei Key Laboratory of Insect Resources Utilization and Sustainable Pest Management, College of Plant Science and Technology, Huazhong Agricultural University, Wuhan 430070, China.
Institute for Advanced Study, Shenzhen University, Shenzhen 518060, China.
email: haominan@szu.edu.cn, ahm1994@126.com
Funding information
This work was supported by National Natural Science Foundation of China (NSFC) grants (32172391). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.
Dryad dataset
Dataset DOI: 10.5061/dryad.79cnp5j7c
Description of the data and file structure
We used dsRNA to knock down the key lysine acetyltransferase p300/CBP gene in the cabbage beetle (Colaphellus bowringi), a Coleopteran vegetable pest that undergoes reproductive diapause induced by a long photoperiod. By measuring ovarian phenotypes, lipid accumulation levels, juvenile hormone (JH) synthesis, and JH response levels, we analyzed the mechanism by which p300/CBP regulates the occurrence of reproductive diapause in the cabbage beetle through the JH pathway. Additionally, we combined methoprene induction experiments to analyze the specificity of p300/CBP in regulating the ovarian response to JH signaling in the cabbage beetle. SD means short-day, reproductive females. LD means long-day, diapause females. MG means mid-gut, FB means fat bodies, OVA means ovaries. Fig 1c is a schematic diagram of the RNAi target region and contains no actual data; therefore, it is not included in this dataset.
Files and variables
File: IMB_all_data.zip
File names
When applicable, file names follow the pattern
eut.(fig1).R(fig2).(fig3).(fig4).(fig5).(file_extension), where:
(fig1)fig1a_Temporal_expression_pattern.csvgroupfor different group include heads of LD-0dA-head,SD-0dA-head,LD-1dA-head,SD-1dA-head,LD-2dA-head,SD-2dA-head,LD-3dA-head,SD-3dA-head,LD-4dA-head SD-4dA-headbiological replicatefor different biological replicates include 3 biological replicates such as biological replicate 1,biological replicate 2,biological replicate 3technical replicatefor different technical replicates include 2 technical replicates such as technical replicate 1,technical replicate 2ct value of CBPfor ct value of CBPct value of rpl19for ct value of rpl19ct value of act1for ct value of act1Geometric mean of referencesfor mean ct values calculationΔCt (target-reference)for ΔCt values calculationmean ΔCt in technical replicatefor mean ΔCt in technical replicate calculationmean ΔCt in control groupfor mean ΔCt in control group calculationΔΔCt(target gene-reference)for ΔΔCt(target gene-reference) calculation2^-ΔΔCt)for 2^-ΔΔCt calculation
fig1b_Tissue_expression_pattern.CSVgroupfor different group include SD-2dA-MG,SD-2dA-head,SD-2dA-FB,SD-2dA-OVAbiological replicatefor different biological replicates include 3 biological replicates such as biological replicate 1,biological replicate 2,biological replicate 3technical replicatefor different technical replicates include 2 technical replicates such as technical replicate 1,technical replicate 2ct value of CBPfor ct value of CBPct value of rpl19for ct value of rpl19ct value of act1for ct value of act1Geometric mean of referencesfor mean ct values calculationΔCt (target-reference)for ΔCt values calculationmean ΔCt in technical replicatefor mean ΔCt in technical replicate calculationmean ΔCt in control groupfor mean ΔCt in control group calculationΔΔCt(target gene-reference)for ΔΔCt(target gene-reference) calculation2^-ΔΔCt)for 2^-ΔΔCt calculation
fig1d_SD+P-dsCBP_Mortality.csvgroupfor different group include SD+P-dsGFP1,SD+P-dsGFP2,SD+P-dsGFP3,SD+P-dsCBP1,SD+P-dsCBP2,SD+P-dsCBP3death numberfor count death numberMortality %for mortality calculation
fig1e_SD_0dA_dsCBP_Survival_rate.CSVn=30means each group include 30 individualsdays after RNAimeans days after dsRNA injectionSD+dsGFPmeans dsGFP injection controlSD+dsCBPmeans dsCBP injection
fig1f_SD+dsCBP_RNAi_efficiency.CSVgroupfor different group include 4-day-old female adults whole bodies of SD+dsGFP,SD+dsCBPbiological replicatefor different biological replicates include 3 biological replicates such as biological replicate 1,biological replicate 2,biological replicate 3technical replicatefor different technical replicates include 2 technical replicates such as technical replicate 1,technical replicate 2ct value of CBPfor ct value of CBPct value of rpl19for ct value of rpl19ct value of act1for ct value of act1Geometric mean of referencesfor mean ct values calculationΔCt (target-reference)for ΔCt values calculationmean ΔCt in technical replicatefor mean ΔCt in technical replicate calculationmean ΔCt in control groupfor mean ΔCt in control group calculationΔΔCt(target gene-reference)for ΔΔCt(target gene-reference) calculation2^-ΔΔCt)for 2^-ΔΔCt calculation
fig1g_ovary_class.csvgroupfor different group include SD+0dA+dsGFP,SD+0dA+dsCBP-F1,SD+0dA+dsCBP-F2classfor different developmental class of ovaries include class 0,Class I,Class II,Class III,Class IVStandardThe criteria for classifying ovarian stages. Undifferentiated ovarian tubules lacking vitellogenin deposition=class 0,Ovarian tubule enlargement without vitellogenin deposition=Class I,Expanded ovarian tubules containing vitellogenin deposits but lacking defined eggs=Class II,Developed ovarian tubules with vitellogenin deposits and distinct immature eggs (spherical, size-variable precursors)=Class III,Fully matured ovarian tubules packed with uniform oval-shaped eggs=Class IVnumbersnumbers of ovaries of class 0,Class I,Class II,Class III and Class IV%Percentage of class 0,Class I,Class II,Class III oand Class IV
fig1h_SD+dsCBP_ovary_phenotype.zipinclude ovary phenotype pics and ovary size sft format files of SD+0dA+dsGFP,SD+0dA+dsCBP-F1,SD+0dA+dsCBP-F2. Files in sft format can be opened with ScopePhoto. The ScopePhoto software can be downloaded from the address at the end of this document.fig1i_ovary size.csvgroupfor different group include SD+0dA+dsGFP,SD+0dA+dsCBP-F1,SD+0dA+dsCBP-F2figureovary figure name in fig1h_SD+dsCBP_ovary_phenotype.zipLength/PixelsThe pixel values of ovarian length measured using ScopePhoto1mm Scale/PixelsThe pixel values of 1mm ScaleActual LengthThe actual length obtained by dividing Length/Pixels by 1mm Scale/PixelsWidth/PixelsThe pixel values of ovarian width measured using ScopePhotoActual WidthThe actual Width obtained by dividing Width/Pixels by 1mm Scale/Pixels
fig1j_SD+dsCBP_vg1_vg2_RTqPCR.CSVgroupfor different group include 4-day-old female adults fat bodies of SD+dsGFP,SD+dsCBPbiological replicatefor different biological replicates include 3 biological replicates such as biological replicate 1,biological replicate 2,biological replicate 3technical replicatefor different technical replicates include 2 technical replicates such as technical replicate 1,technical replicate 2ct value of vg1for ct value of vg1ct value of vg2for ct value of vg2ct value of rpl19for ct value of rpl19ct value of act1for ct value of act1Geometric mean of referencesfor mean ct values calculationΔCt (target-reference)for ΔCt values calculationmean ΔCt in technical replicatefor mean ΔCt in technical replicate calculationmean ΔCt in control groupfor mean ΔCt in control group calculationΔΔCt(target gene-reference)for ΔΔCt(target gene-reference) calculation2^-ΔΔCt)for 2^-ΔΔCt calculation
fig1k_SD+dsCBP_JHAMT1_HMGR2_FPPS1_RTqPCR.CSVgroupfor different group include 4-day-old female adults heads of SD+dsGFP,SD+dsCBPbiological replicatefor different biological replicates include 3 biological replicates such as biological replicate 1,biological replicate 2,biological replicate 3technical replicatefor different technical replicates include 2 technical replicates such as technical replicate 1,technical replicate 2ct value of JHAMT1for ct value of JHAMT1ct value of HMGR2for ct value of HMGR2ct value of FPPS1for ct value of FPPS1ct value of rpl19for ct value of rpl19ct value of act1for ct value of act1Geometric mean of referencesfor mean ct values calculationΔCt (target-reference)for ΔCt values calculationmean ΔCt in technical replicatefor mean ΔCt in technical replicate calculationmean ΔCt in control groupfor mean ΔCt in control group calculationΔΔCt(target gene-reference)for ΔΔCt(target gene-reference) calculation2^-ΔΔCt)for 2^-ΔΔCt calculation
fig1l_SD+dsCBP_JHAMT1_WB.jpginclude WB data of JHAMT1 in 2-day-old female adults heads of SD+dsGFP and SD+dsCBPfig1m_SD+dsCBP_JHAMT1_relative_protein_level.CSVgroupfor different group include -day-old female adults heads SD+dsGFP1,SD+dsGFP2,SD+dsGFP3,SD+dsCBP1,SD+dsCBP2,SD+dsCBP3JHAMT1values of JHAMT1 proteinAverage value of JHAMT1 in GFPaverage value calculation of JHAMT1 protein valueRelative JHAMT1caculation of relative JHAMT1 protein levelα Tublinvalues of α Tublin proteinJAverage value of α Tublin in GFPaverage value calculation of α Tublin protein valueRelative α Tublincaculation of relative α Tublin protein levelJHAMT1/α Tublincaculation of relative JHAMT1 protein level to α Tublin protein level
fig1n_SD+dsCBP_HPLC_MSMS.CSVHemolymph volumehemolymph volume from 4-day-old female adults of dsGFP and dsCBP is 50μLConcentrated volumevolume of sample after extraction, concentration, and reconstitutionConcentration factorconcentration factor (fold) of the sample during preparation of the final assay solutionJH III (ng/mL)=(Peak area+379686)/25553/Concentration factorJH titer Calculation Formulagroupfor different group include SD+dsGFP1,SD+dsGFP2,SD+dsGFP3,SD+dsCBP1,SD+dsCBP2,SD+dsCBP3Peak areaaverage value calculation of α Tublin protein valueJH III (ng/mL)caculation of relative α Tublin protein level
fig1n_SD+dsCBP_HPLC_MSMS.jpginclude HPLC/MSMS data of standard JHIII and samples of SD+dsGFP and SD+dsCBPfig1o_SD+dsCBP_krh1_JHE1_RTqPCR.CSVgroupfor different group include 4-day-old reproductive female adults fat bodies of SD+dsGFP,SD+dsCBPbiological replicatefor different biological replicates include 3 biological replicates such as biological replicate 1,biological replicate 2,biological replicate 3technical replicatefor different technical replicates include 2 technical replicates such as technical replicate 1,technical replicate 2ct value of krh1for ct value of kr-h1ct value of JHE1for ct value of JHE1ct value of rpl19for ct value of rpl19ct value of act1for ct value of act1Geometric mean of referencesfor mean ct values calculationΔCt (target-reference)for ΔCt values calculationmean ΔCt in technical replicatefor mean ΔCt in technical replicate calculationmean ΔCt in control groupfor mean ΔCt in control group calculationΔΔCt(target gene-reference)for ΔΔCt(target gene-reference) calculation2^-ΔΔCt)for 2^-ΔΔCt calculation
(fig2)fig2a_SD+dsCBP_Nile_red.jpginclude nile red data of 4-day-old reproductive female adults fat bodies of SD+dsGFP and SD+dsCBPfig2b_SD+dsCBP_Nile_red_measurement.CSVSD+dsGFPgroup of SD+dsGFP include 40 lipid dropletsSD+dsCBPgroup of SD+dsCBP include 40 lipid dropletsscale/μmphoto scale is 100μmpixelpixel value corresponding to the scale barlength 1measurement 1length 2measurement 2averageaverage of the two measurementsactual length/μmcaculation of actual length
fig2b_SD+dsCBP_Nile_red_measurement.jpginclude nile red measurement picfig2c_SD+dsCBP_TG.CSVGroupgroup of Ethanol(blank),1/3 standard,standard,4-day-old reproductive female adults whole bodies of SD-dsGFP-4d-1,SD-dsGFP-4d-2,SD-dsGFP-4d-3,SD-dsGFP-4d-4,SD-dsGFP-4d-5,SD-dsCBP-4d-1,SD-dsCBP-4d-2,SD-dsCBP-4d-3,SD-dsCBP-4d-4,SD-dsCBP-4d-5OD510OD510 valueaverage ODaverage OD510sample OD-blank ODsample OD-blank ODstandard OD-blank ODstandard OD-blank OD(sample OD-blank OD)/(standard OD-blank OD)(sample OD-blank OD)/(standard OD-blank OD)Standard concentration(mmol/L)Standard concentration(mmol/L)[(sample OD-blank OD)/(standard OD-blank OD)]*Standard concentration[(sample OD-blank OD)/(standard OD-blank OD)]*Standard concentrationFresh weight(gFw)Fresh weight value of each samplesample TG concentration(gFW/L)sample TG concentration(gFW/L)TG(mmol/gFw)TG valueaverage of SD-dsGFPaverage TG valueRelative TG=TG/(Average of SD-dsGFP)relative TG Calculation Formula
fig2d_SD+dsCBP_Total_lipid.CSVGroupgroup of 4-day-old reproductive female adults whole bodies of SD+dsGFP-1 to SD+dsGFP-10,SD+dsCBP-1 to SD+dsCBP-10Weight of the glass weighing bottle(W/g)Weight of the glass weighing bottle(W/g)initial dry weight(IDW/g)initial dry weight(IDW/g)dry weight after extration(EDW/g)dry weight after extration(EDW/g)TL%=(IDW-EDW)/(IDW-W)×100 %TL Calculation Formula
fig2e_SD+dsCBP_HSP21_23_70_RTqPCR.CSVgroupfor different group include 4-day-old reproductive female adults whole bodies of SD+dsGFP,SD+dsCBPbiological replicatefor different biological replicates include 3 biological replicates such as biological replicate 1,biological replicate 2,biological replicate 3technical replicatefor different technical replicates include 2 technical replicates such as technical replicate 1,technical replicate 2ct value of HSP21for ct value of HSP21ct value of HSP23for ct value of HSP23ct value of HSP70for ct value of HSP70ct value of rpl19for ct value of rpl19ct value of act1for ct value of act1Geometric mean of referencesfor mean ct values calculationΔCt (target-reference)for ΔCt values calculationmean ΔCt in technical replicatefor mean ΔCt in technical replicate calculationmean ΔCt in control groupfor mean ΔCt in control group calculationΔΔCt(target gene-reference)for ΔΔCt(target gene-reference) calculation2^-ΔΔCt)for 2^-ΔΔCt calculation
fig2f_SD+dsCBP_DP1_RTqPCR.CSVgroupfor different group include 4-day-old reproductive female adults fat bodies of SD+dsGFP,SD+dsCBPbiological replicatefor different biological replicates include 3 biological replicates such as biological replicate 1,biological replicate 2,biological replicate 3technical replicatefor different technical replicates include 2 technical replicates such as technical replicate 1,technical replicate 2ct value of DP1for ct value of DP1ct value of rpl19for ct value of rpl19ct value of act1for ct value of act1Geometric mean of referencesfor mean ct values calculationΔCt (target-reference)for ΔCt values calculationmean ΔCt in technical replicatefor mean ΔCt in technical replicate calculationmean ΔCt in control groupfor mean ΔCt in control group calculationΔΔCt(target gene-reference)for ΔΔCt(target gene-reference) calculation2^-ΔΔCt)for 2^-ΔΔCt calculation
(fig3)fig3aSDi+ME_ovary_phenotype.zipinclude ovary phenotype pics and ovary size sft format files of 4-day-old reproductive female adults of SD+dsGFP+Ace,SD+dsCBP+Ace,SD+dsCBP+ME. Files in sft format can be opened with ScopePhoto. The ScopePhoto software can be downloaded from the address at the end of this document.fig3b_ovary_class.csvgroupfor different group include SD+dsGFP+Ace,SD+dsCBP+Ace,SD+dsCBP+MEclassfor different developmental class of ovaries include class 0,Class I,Class II,Class III,Class IVStandardThe criteria for classifying ovarian stages. Undifferentiated ovarian tubules lacking vitellogenin deposition=class 0,Ovarian tubule enlargement without vitellogenin deposition=Class I,Expanded ovarian tubules containing vitellogenin deposits but lacking defined eggs=Class II,Developed ovarian tubules with vitellogenin deposits and distinct immature eggs (spherical, size-variable precursors)=Class III,Fully matured ovarian tubules packed with uniform oval-shaped eggs=Class IVnumbersnumbers of ovaries of class 0,Class I,Class II,Class III and Class IV%Percentage of class 0,Class I,Class II,Class III oand Class IV
fig3c_ovary_size.csvgroupfor different group include SD+dsGFP+Ace,SD+dsCBP+Ace,SD+dsCBP+MEfigureovary figure name in fig1h_SD+dsCBP_ovary_phenotype.zipLength/PixelsThe pixel values of ovarian length measured using ScopePhoto1mm Scale/PixelsThe pixel values of 1mm ScaleActual LengthThe actual length obtained by dividing Length/Pixels by 1mm Scale/PixelsWidth/PixelsThe pixel values of ovarian width measured using ScopePhotoActual WidthThe actual Width obtained by dividing Width/Pixels by 1mm Scale/Pixels
fig3d_SDi+ME_RNAi_efficiency.CSVgroupfor different group include 4-day-old reproductive female adults whole bodies of SD+dsGFP+Ace,SD+dsCBP+Ace,SD+dsCBP+MEbiological replicatefor different biological replicates include 3 biological replicates such as biological replicate 1,biological replicate 2,biological replicate 3technical replicatefor different technical replicates include 2 technical replicates such as technical replicate 1,technical replicate 2ct value of CBPfor ct value of CBPct value of rpl19for ct value of rpl19ct value of act1for ct value of act1Geometric mean of referencesfor mean ct values calculationΔCt (target-reference)for ΔCt values calculationmean ΔCt in technical replicatefor mean ΔCt in technical replicate calculationmean ΔCt in control groupfor mean ΔCt in control group calculationΔΔCt(target gene-reference)for ΔΔCt(target gene-reference) calculation2^-ΔΔCt)for 2^-ΔΔCt calculation
fig3e_SDi+ME_JHE1_krh1_RTqPCR.CSVgroupfor different group include 4-day-old reproductive female adults fat bodies of SD+dsGFP+Ace,SD+dsCBP+Ace,SD+dsCBP+MEbiological replicatefor different biological replicates include 3 biological replicates such as biological replicate 1,biological replicate 2,biological replicate 3technical replicatefor different technical replicates include 2 technical replicates such as technical replicate 1,technical replicate 2ct value of krh1for ct value of kr-h1ct value of JHE1for ct value of JHE1ct value of rpl19for ct value of rpl19ct value of act1for ct value of act1Geometric mean of referencesfor mean ct values calculationΔCt (target-reference)for ΔCt values calculationmean ΔCt in technical replicatefor mean ΔCt in technical replicate calculationmean ΔCt in control groupfor mean ΔCt in control group calculationΔΔCt(target gene-reference)for ΔΔCt(target gene-reference) calculation2^-ΔΔCt)for 2^-ΔΔCt calculation
fig3f_SDi+ME_vg1_vg2_RTqPCR.CSVgroupfor different group include 4-day-old reproductive female adults fat bodies of SD+dsGFP+Ace,SD+dsCBP+Ace,SD+dsCBP+MEbiological replicatefor different biological replicates include 3 biological replicates such as biological replicate 1,biological replicate 2,biological replicate 3technical replicatefor different technical replicates include 2 technical replicates such as technical replicate 1,technical replicate 2ct value of vg1for ct value of vg1ct value of vg2for ct value of vg2ct value of rpl19for ct value of rpl19ct value of act1for ct value of act1Geometric mean of referencesfor mean ct values calculationΔCt (target-reference)for ΔCt values calculationmean ΔCt in technical replicatefor mean ΔCt in technical replicate calculationmean ΔCt in control groupfor mean ΔCt in control group calculationΔΔCt(target gene-reference)for ΔΔCt(target gene-reference) calculation2^-ΔΔCt)for 2^-ΔΔCt calculation
fig3g_SDi+ME_Nile_red.jpginclude nile red data of fat bodies of 4-day-old reproductive female adults of SD+dsGFP+Ace,SD+dsCBP+Ace,SD+dsCBP+MEfig3h_SDi+ME_Nile_red_measurement.CSVSD+dsGFP+Acegroup of 4-day-old reproductive female adults fat bodies of SD+dsGFP+Ace include 40 lipid dropletsSD+dsCBP+Acegroup of 4-day-old reproductive female adults fat bodies of SD+dsCBP+Ace include 40 lipid dropletsSD+dsCBP+MEgroup of 4-day-old reproductive female adults fat bodies of SD+dsCBP+ME include 40 lipid dropletsscale/μmphoto scale is 100μmpixelpixel value corresponding to the scale barlength 1measurement 1length 2measurement 2averageaverage of the two measurementsactual length/μmcaculation of actual length
fig3h_SDi+ME_Nile_red_measurement.jpginclude nile red measurement picfig3i_SDi+ME_TG.CSVGroupgroup of Ethanol(blank),1/3 standard,standard,4-day-old reproductive female adults whole bodies of SD-dsGFP-Ace-1,SSD-dsGFP-Ace-2,SD-dsGFP-Ace-3,SD-dsCBP-Ace-1,SD-dsCBP-Ace-2,SD-dsCBP-Ace-3,SD-dsCBP-ME-1,SD-dsCBP-ME-2,SD-dsCBP-ME-3average ODaverage OD510sample OD-blank ODsample OD-blank ODstandard OD-blank ODstandard OD-blank OD(sample OD-blank OD)/(standard OD-blank OD)(sample OD-blank OD)/(standard OD-blank OD)Standard concentration(mmol/L)Standard concentration(mmol/L)[(sample OD-blank OD)/(standard OD-blank OD)]*Standard concentration[(sample OD-blank OD)/(standard OD-blank OD)]*Standard concentrationFresh weight(gFw)Fresh weight value of each samplesample TG concentration(gFW/L)sample TG concentration(gFW/L)TG(mmol/gFw)TG valueaverage of SD-dsGFP-Aceaverage TG valueRelative TG=TG/(Average of SD-dsGFP-Ace)relative TG Calculation Formula
fig3j_SDi+ME_total_lipid.CSVGroupgroup of 4-day-old reproductive female adults whole bodies of SD+dsGFP+Ace-1 to SD+dsGFP+Ace-13,SD+dsCBP+Ace-1 to SD+dsCBP+Ace-13,SD+dsCBP+ME-1 to SD+dsCBP+ME-13Weight of the glass weighing bottle(W/g)Weight of the glass weighing bottle(W/g)initial dry weight(IDW/g)initial dry weight(IDW/g)dry weight after extration(EDW/g)dry weight after extration(EDW/g)TL%=(IDW-EDW)/(IDW-W)×100 %TL Calculation Formula
(fig4)fig4a_LDi+ME_ovary_phenotype.zipinclude ovary phenotype pics and ovary size sft format files of 4-day-old diapause female adults of LD+dsGFP+Ace,LD+dsGFP+ME,LD+dsCBP+Ace,LD+dsCBP+ME. Files in sft format can be opened with ScopePhoto. The ScopePhoto software can be downloaded from the address at the end of this document.fig4b_ovary_class.csvgroupfor different group include LD+dsGFP+Ace,LD+dsGFP+ME,LD+dsCBP+Ace,LD+dsCBP+MEclassfor different developmental class of ovaries include class 0,Class I,Class II,Class III,Class IVStandardThe criteria for classifying ovarian stages. Undifferentiated ovarian tubules lacking vitellogenin deposition=class 0,Ovarian tubule enlargement without vitellogenin deposition=Class I,Expanded ovarian tubules containing vitellogenin deposits but lacking defined eggs=Class II,Developed ovarian tubules with vitellogenin deposits and distinct immature eggs (spherical, size-variable precursors)=Class III,Fully matured ovarian tubules packed with uniform oval-shaped eggs=Class IVnumbersnumbers of ovaries of class 0,Class I,Class II,Class III and Class IV%Percentage of class 0,Class I,Class II,Class III oand Class IV
fig4c_ovary_size.csvgroupfor different group include LD+dsGFP+Ace,LD+dsGFP+ME,LD+dsCBP+Ace,LD+dsCBP+MEfigureovary figure name in fig1h_SD+dsCBP_ovary_phenotype.zipLength/PixelsThe pixel values of ovarian length measured using ScopePhoto1mm Scale/PixelsThe pixel values of 1mm ScaleActual LengthThe actual length obtained by dividing Length/Pixels by 1mm Scale/PixelsWidth/PixelsThe pixel values of ovarian width measured using ScopePhotoActual WidthThe actual Width obtained by dividing Width/Pixels by 1mm Scale/Pixels
fig4d_LDi+ME RNAi_efficiency_RTqPCR.CSVgroupfor different group include 4-day-old reproductive female adults whole bodies of LD+dsGFP+Ace,LD+dsGFP+ME,LD+dsCBP+Ace,LD+dsCBP+MEbiological replicatefor different biological replicates include 3 biological replicates such as biological replicate 1,biological replicate 2,biological replicate 3technical replicatefor different technical replicates include 2 technical replicates such as technical replicate 1,technical replicate 2ct value of CBPfor ct value of CBPct value of rpl19for ct value of rpl19ct value of act1for ct value of act1Geometric mean of referencesfor mean ct values calculationΔCt (target-reference)for ΔCt values calculationmean ΔCt in technical replicatefor mean ΔCt in technical replicate calculationmean ΔCt in control groupfor mean ΔCt in control group calculationΔΔCt(target gene-reference)for ΔΔCt(target gene-reference) calculation2^-ΔΔCt)for 2^-ΔΔCt calculation
fig4e_LDi+ME_Vg1_Vg2_RTqPCR.CSVgroupfor different group include 4-day-old reproductive female adults fat bodies of LD+dsGFP+Ace,LD+dsGFP+ME,LD+dsCBP+Ace,LD+dsCBP+MEbiological replicatefor different biological replicates include 3 biological replicates such as biological replicate 1,biological replicate 2,biological replicate 3technical replicatefor different technical replicates include 2 technical replicates such as technical replicate 1,technical replicate 2ct value of vg1for ct value of vg1ct value of vg2for ct value of vg2ct value of rpl19for ct value of rpl19ct value of act1for ct value of act1Geometric mean of referencesfor mean ct values calculationΔCt (target-reference)for ΔCt values calculationmean ΔCt in technical replicatefor mean ΔCt in technical replicate calculationmean ΔCt in control groupfor mean ΔCt in control group calculationΔΔCt(target gene-reference)for ΔΔCt(target gene-reference) calculation2^-ΔΔCt)for 2^-ΔΔCt calculation
fig4f_LDi+ME_krh1_JHE1_RTqPCR.CSVgroupfor different group include 4-day-old reproductive female adults fat bodies of LD+dsGFP+Ace,LD+dsGFP+ME,LD+dsCBP+Ace,LD+dsCBP+MEbiological replicatefor different biological replicates include 3 biological replicates such as biological replicate 1,biological replicate 2,biological replicate 3technical replicatefor different technical replicates include 2 technical replicates such as technical replicate 1,technical replicate 2ct value of krh1for ct value of kr-h1ct value of JHE1for ct value of JHE1ct value of rpl19for ct value of rpl19ct value of act1for ct value of act1Geometric mean of referencesfor mean ct values calculationΔCt (target-reference)for ΔCt values calculationmean ΔCt in technical replicatefor mean ΔCt in technical replicate calculationmean ΔCt in control groupfor mean ΔCt in control group calculationΔΔCt(target gene-reference)for ΔΔCt(target gene-reference) calculation2^-ΔΔCt)for 2^-ΔΔCt calculation
fig4g_LDi+ME_Nile_red.jpginclude nile red data of fat bodies of 4-day-old reproductive female adults of LD+dsGFP+Ace,LD+dsGFP+ME,LD+dsCBP+Ace,LD+dsCBP+MEfig4h_LDi+ME_Nile_Red_measurement.CSVLD+dsGFP+Acegroup of LD+dsGFP+Ace include 40 lipid dropletsLD+dsGFP+MEgroup of LD+dsGFP+ME include 40 lipid dropletsLD+dsCBP+Acegroup of LD+dsCBP+Ace include 40 lipid dropletsLD+dsCBP+MEgroup of LD+dsCBP+ME include 40 lipid dropletsscale/μmphoto scale is 100μmpixelpixel value corresponding to the scale barlength 1measurement 1length 2measurement 2averageaverage of the two measurementsactual length/μmcaculation of actual length
fig4h_LDi+ME_Nile_Red_measurement.jpginclude nile red measurement picfig4i_LDi+ME_TG.CSVGroupgroup of Ethanol(blank),1/3 standard,standard,4-day-old reproductive female adults whole bodies of LD-dsGFP-Ace-1,LD-dsGFP-Ace-2,LD-dsGFP-Ace-3,LD-dsGFP-Ace-4,LD-dsGFP-ME-1,LD-dsGFP-ME-2,LD-dsGFP-ME-3,LD-dsGFP-ME-4,LD-dsCBP-Ace-1,LD-dsCBP-Ace-2,LD-dsCBP-Ace-3,LD-dsCBP-Ace-4,LD-dsCBP-ME-1,LD-dsCBP-ME-2,LD-dsCBP-ME-3,LD-dsCBP-ME-4average ODaverage OD510average ODaverage OD510sample OD-blank ODsample OD-blank ODstandard OD-blank ODstandard OD-blank OD(sample OD-blank OD)/(standard OD-blank OD)(sample OD-blank OD)/(standard OD-blank OD)Standard concentration(mmol/L)Standard concentration(mmol/L)[(sample OD-blank OD)/(standard OD-blank OD)]*Standard concentration[(sample OD-blank OD)/(standard OD-blank OD)]*Standard concentrationFresh weight(gFw)Fresh weight value of each samplesample TG concentration(gFW/L)sample TG concentration(gFW/L)TG(mmol/gFw)TG valueaverage of LD+dsGFP+Aceaverage TG valueRelative TG=TG/(Average of LD+dsGFP+Ace)relative TG Calculation Formula
fig4j_LDi+ME_total_lipid.CSVGroupgroup of 4-day-old reproductive female adults whole bodies of LD+dsGFP+Ace-1 to LD+dsGFP+Ace-13,LD+dsGFP+ME-1 to LD+dsGFP+ME-13,LD+dsCBP+Ace-1 to LD+dsCBP+Ace-13,LD+dsCBP+ME-1 to LD+dsCBP+ME-13Weight of the glass weighing bottle(W/g)Weight of the glass weighing bottle(W/g)initial dry weight(IDW/g)initial dry weight(IDW/g)dry weight after extration(EDW/g)dry weight after extration(EDW/g)TL%=(IDW-EDW)/(IDW-W)×100 %TL Calculation Formula
(fig5)fig5a_SDi+ME_TGL1_RTqPCR.CSVgroupfor different group include 4-day-old reproductive female adults fat bodies of SD+dsGFP+Ace,SD+dsCBP+Ace,SD+dsCBP+MEbiological replicatefor different biological replicates include 3 biological replicates such as biological replicate 1,biological replicate 2,biological replicate 3technical replicatefor different technical replicates include 2 technical replicates such as technical replicate 1,technical replicate 2ct value of TGL1for ct value of TGL1ct value of rpl19for ct value of rpl19ct value of act1for ct value of act1Geometric mean of referencesfor mean ct values calculationΔCt (target-reference)for ΔCt values calculationmean ΔCt in technical replicatefor mean ΔCt in technical replicate calculationmean ΔCt in control groupfor mean ΔCt in control group calculationΔΔCt(target gene-reference)for ΔΔCt(target gene-reference) calculation2^-ΔΔCt)for 2^-ΔΔCt calculation
fig5b_LDi+ME_TGL1_RTqPCR.CSVgroupfor different group include 4-day-old diapause female adults fat bodies of LD+dsGFP+Ace,LD+dsGFP+ME,LD+dsCBP+Ace,LD+dsCBP+MEbiological replicatefor different biological replicates include 3 biological replicates such as biological replicate 1,biological replicate 2,biological replicate 3technical replicatefor different technical replicates include 2 technical replicates such as technical replicate 1,technical replicate 2ct value of TGL1for ct value of TGL1ct value of rpl19for ct value of rpl19ct value of act1for ct value of act1Geometric mean of referencesfor mean ct values calculationΔCt (target-reference)for ΔCt values calculationmean ΔCt in technical replicatefor mean ΔCt in technical replicate calculationmean ΔCt in control groupfor mean ΔCt in control group calculationΔΔCt(target gene-reference)for ΔΔCt(target gene-reference) calculation2^-ΔΔCt)for 2^-ΔΔCt calculation
fig5c_SDi+ME_krh1_RTqPCR.CSVgroupfor different group include 4-day-old reproductive female adults fat bodies of SD+dsGFP+Ace,SD+dsCBP+Ace,SD+dsCBP+MEbiological replicatefor different biological replicates include 3 biological replicates such as biological replicate 1,biological replicate 2,biological replicate 3technical replicatefor different technical replicates include 2 technical replicates such as technical replicate 1,technical replicate 2ct value of krh1for ct value of Kr-h1ct value of rpl19for ct value of rpl19ct value of act1for ct value of act1Geometric mean of referencesfor mean ct values calculationΔCt (target-reference)for ΔCt values calculationmean ΔCt in technical replicatefor mean ΔCt in technical replicate calculationmean ΔCt in control groupfor mean ΔCt in control group calculationΔΔCt(target gene-reference)for ΔΔCt(target gene-reference) calculation2^-ΔΔCt)for 2^-ΔΔCt calculation
fig5d_SDi+ME_Mcm4_RTqPCR.CSVgroupfor different group include 4-day-old reproductive female adults fat bodies of SD+dsGFP+Ace,SD+dsCBP+Ace,SD+dsCBP+MEbiological replicatefor different biological replicates include 3 biological replicates such as biological replicate 1,biological replicate 2,biological replicate 3technical replicatefor different technical replicates include 2 technical replicates such as technical replicate 1,technical replicate 2ct value of Mcm4for ct value of Mcm4ct value of rpl19for ct value of rpl19ct value of act1for ct value of act1Geometric mean of referencesfor mean ct values calculationΔCt (target-reference)for ΔCt values calculationmean ΔCt in technical replicatefor mean ΔCt in technical replicate calculationmean ΔCt in control groupfor mean ΔCt in control group calculationΔΔCt(target gene-reference)for ΔΔCt(target gene-reference) calculation2^-ΔΔCt)for 2^-ΔΔCt calculation
fig5e_LDi+ME_krh1_RTqPCR.CSVgroupfor different group include 4-day-old diapause female adults fat bodies of LD+dsGFP+Ace,LD+dsGFP+ME,LD+dsCBP+Ace,LD+dsCBP+MEbiological replicatefor different biological replicates include 3 biological replicates such as biological replicate 1,biological replicate 2,biological replicate 3technical replicatefor different technical replicates include 2 technical replicates such as technical replicate 1,technical replicate 2ct value of krh1for ct value of Kr-h1ct value of rpl19for ct value of rpl19ct value of act1for ct value of act1Geometric mean of referencesfor mean ct values calculationΔCt (target-reference)for ΔCt values calculationmean ΔCt in technical replicatefor mean ΔCt in technical replicate calculationmean ΔCt in control groupfor mean ΔCt in control group calculationΔΔCt(target gene-reference)for ΔΔCt(target gene-reference) calculation2^-ΔΔCt)for 2^-ΔΔCt calculation
fig5f_LDi+ME_Mcm4_RTqPCR.CSVgroupfor different group include 4-day-old diapause female adults fat bodies of LD+dsGFP+Ace,LD+dsGFP+ME,LD+dsCBP+Ace,LD+dsCBP+MEbiological replicatefor different biological replicates include 3 biological replicates such as biological replicate 1,biological replicate 2,biological replicate 3technical replicatefor different technical replicates include 2 technical replicates such as technical replicate 1,technical replicate 2ct value of Mcm4for ct value of Mcm4ct value of rpl19for ct value of rpl19ct value of act1for ct value of act1Geometric mean of referencesfor mean ct values calculationΔCt (target-reference)for ΔCt values calculationmean ΔCt in technical replicatefor mean ΔCt in technical replicate calculationmean ΔCt in control groupfor mean ΔCt in control group calculationΔΔCt(target gene-reference)for ΔΔCt(target gene-reference) calculation2^-ΔΔCt)for 2^-ΔΔCt calculation
fig5g_SDi+ME_VgR_RTqPCR.CSVgroupfor different group include 4-day-old reproductive female adults fat bodies of SD+dsGFP+Ace,SD+dsCBP+Ace,SD+dsCBP+MEbiological replicatefor different biological replicates include 3 biological replicates such as biological replicate 1,biological replicate 2,biological replicate 3technical replicatefor different technical replicates include 2 technical replicates such as technical replicate 1,technical replicate 2ct value of VgRfor ct value of VgRct value of rpl19for ct value of rpl19ct value of act1for ct value of act1Geometric mean of referencesfor mean ct values calculationΔCt (target-reference)for ΔCt values calculationmean ΔCt in technical replicatefor mean ΔCt in technical replicate calculationmean ΔCt in control groupfor mean ΔCt in control group calculationΔΔCt(target gene-reference)for ΔΔCt(target gene-reference) calculation2^-ΔΔCt)for 2^-ΔΔCt calculation
fig5h_LDi+ME_VgR_RTqPCR.CSVgroupfor different group include 4-day-old diapause female adults fat bodies of LD+dsGFP+Ace,LD+dsGFP+ME,LD+dsCBP+Ace,LD+dsCBP+MEbiological replicatefor different biological replicates include 3 biological replicates such as biological replicate 1,biological replicate 2,biological replicate 3technical replicatefor different technical replicates include 2 technical replicates such as technical replicate 1,technical replicate 2ct value of VgRfor ct value of VgRct value of rpl19for ct value of rpl19ct value of act1for ct value of act1Geometric mean of referencesfor mean ct values calculationΔCt (target-reference)for ΔCt values calculationmean ΔCt in technical replicatefor mean ΔCt in technical replicate calculationmean ΔCt in control groupfor mean ΔCt in control group calculationΔΔCt(target gene-reference)for ΔΔCt(target gene-reference) calculation2^-ΔΔCt)for 2^-ΔΔCt calculation
Description:
The data has been organized in the order of the figures.
Code/software
Scopephoto was used to measure the size of the ovaries or lipid droplets.
