General information

Related article:
Wu Y-K, Yu J-Q, Jin L, Li G-Q. A basic helix-loop-helix/Per-Arnt-Sim transcription factor Taiman is essential for metamorphosis in Henosepilachna vigintioctomaculata. Insect Molecular Biology, forthcoming.
Species: Henosepilachna vigintioctomaculata
Main techniques: RNA interference (RNAi), qRT-PCR, body weight measurement, survival analysis.

All data in this repository correspond to Figures 1–5 in the associated article.

File list

rawdata.xlsx

Excel workbook containing all numerical data used to generate Figures 1–5.

No image files or narrative text from the article are included here. All values are numeric and intended for re-analysis.

Conventions and units

• Time after dsRNA injection is given in:
  • hours (h) for short-term measurements (e.g., 0 h, 24 h, …);
  • days (d) for longer observations (e.g,. day 1, day 2, …).
• Body weight is expressed in milligrams (mg).
• qRT-PCR data are given as normalized relative expression values (2^{-ΔCt or 2}-ΔΔCt, unitless).
• For qRT-PCR tables:
  • the first column in each block is the sample label (stage, tissue, or treatment);
  • the next three columns are biological replicates of the normalized expression value
    (three independent pools of individuals, each measured once).

GraphPad Prism output rows (e.g., “Paired t test”, “P value”, “Multiple t test”) are kept in some sheets as summary statistics and can be ignored if only raw measurements are needed.

Detailed description of rawdata.xlsx

Workbook: rawdata.xlsx
Sheets: “Fig1”, “Fig2”, “Fig3”, “Fig4”, “Fig5”.

All row and column references below use Excel conventions (columns A, B, C…).

Sheet “Fig1” – HvTai expression (Figure 1)

This sheet contains two qRT-PCR tables for HvTai:

(1) Figure 1A – Temporal expression of HvTai

• Data block: rows 3–26, columns C–F.
• Row 3 (header):

• C3: “HvmTai” (gene name)
• D3–F3: “2-△ct” (three biological replicates; unitless relative expression)

• Rows 4–26:

• Column C: developmental stage / time point label
  (e.g. “L1D0”, “L1D1”, …, “PD4”).
• Columns D–F: three biological replicates of HvTai relative expression
  (2^-ΔCt) for each stage.

(2) Figure 1B – Tissue expression of HvTai

• Data block: rows 3–10, columns Q–T.
• Row 3 (header):

• Q3: “HvmTai”
• R3–T3: “2-△ct” (three biological replicates; unitless)

• Rows 4–10:

• Column Q: tissue name
  (Head, Foregut, Midgut, Hindgut, Malpighian tubules, Fat body, Epidermis).
• Columns R–T: three biological replicates of HvTai relative expression
  (2^-ΔCt) in each tissue.

There are no separate columns for raw Ct values in this sheet; only normalized 2^-ΔCt values are provided.

Sheet “Fig2” – RNAi at second instar (Figure 2)

This sheet contains:

(1) Figure 2A – Knockdown efficiency of HvTai

• Data block: rows 3–5, columns C–F.
• Row 3:

• C3: “HvmTai”
• D3–F3: “2-△△ct” (three biological replicates; unitless)

• Rows 4–5:

• Column C: treatment group (“dsegfp” control or “dsTai”).
• Columns D–F: three biological replicates of HvTai relative expression
  (2^-ΔΔCt) in each treatment.

(2) Figure 2B – Fresh weight of second-instar larvae after dsRNA injection

• Data block: rows 4–10, columns L–AF.
• Row 3:

• L3: “Fresh weight (mg)”
• M3: “dsegfp”
• W3: “dsTai”

• Rows 4–10:

• Column L: time after injection (0 h, 24 h, 48 h, 72 h, 96 h, 120 h, 144 h).
• Columns M–V: fresh weights (mg) of 10 individual larvae in the dsegfp control group.
• Columns W–AF: fresh weights (mg) of 10 individual larvae in the dsTai-treated group.

(3) Additional rows for Figure 2B (“Paired t test”, “P value”, “Multiple t test”)

• Rows 8–25 contain GraphPad Prism output for statistical comparisons
  between dsegfp and dsTai body weights at each time point
  (P values, significance, t ratio, degrees of freedom, etc.).
• These rows are derived statistics and not raw measurements.

(4) Figure 2C–2D – Survival and developmental stage distribution

• Data block: starting around row 39, columns C–Y.
• Row 40 (approximate; the exact row is labelled “Days”):

• Column C: “Days” (days after injection).
• Columns D and E: numbers or proportions of surviving larvae
  in the dsegfp and dsTai groups (“dsegfp”, “dsTai”).
• Columns with headers such as “Normal pupa”, “Deformed larva”, “Deformed pupa”
  contain the numbers of individuals in each phenotype category on each day.

These data correspond to the Kaplan–Meier survival curves and phenotypic ratios
shown in Figure 2C and 2D.

Sheet “Fig3” – RNAi at third instar (Figure 3)

Structure is similar to “Fig2”:

(1) Figure 3A – Knockdown efficiency of HvTai

• Data block: rows 3–5, columns C–F.
• Row 3:

• C3: “HvmTai”
• D3–F3: “2-△△ct”

• Rows 4–5:

• Column C: treatment group (“dsegfp”, “dsTai”).
• Columns D–F: three biological replicates of HvTai relative expression (2^-ΔΔCt).

(2) Figure 3B – Fresh weight of third-instar larvae

• Data block: rows 4–15, columns L–AF.
• Row 3:

• L3: “Fresh weight (mg)”
• M3: “dsegfp”
• W3: “dsTai”

• Rows 4–15:

• Column L: time after injection (days 1–12).
• Columns M–V: fresh weights (mg) of 10 dsegfp control larvae.
• Columns W–AF: fresh weights (mg) of 10 dsTai-injected larvae.

• Subsequent rows store GraphPad Prism “Multiple t test” outputs
  for comparisons between treatments (P value, mean difference, etc.).

(3) Figure 3C–3D – Survival and phenotypes

• Data block: as in “Fig2”, a table with:

• Column labelled “Days”: days after injection.
• Columns “dsegfp” and “dsTai”: survivors in each group.
• Additional phenotype columns for different developmental outcomes
  (e.g., “Normal pupa”, “Deformed prepupa/pupa”), used to generate Figure 3C–3D.

Sheet “Fig4” – RNAi at fourth instar (Figure 4)

(1) Figure 4A – Knockdown efficiency of HvTai

• Data block: rows 3–5, columns C–F.
• Row 3:

• C3: “HvmTai”
• D3–F3: “2-△△ct”

• Rows 4–5:

• Column C: treatment group (“dsegfp”, “dsTai”).
• Columns D–F: three biological replicates of HvTai relative expression (2^-ΔΔCt).

(2) Figure 4B – Fresh weight of fourth-instar larvae

• Data block: rows 4–13, columns L–AF.
• Row 3:

• L3: “Fresh weight (mg)”
• M3: “dsegfp”
• W3: “dsTai”

• Rows 4–13:

• Column L: time after injection (0 h, 24 h, 48 h, 72 h, 96 h, 120 h, 144 h, 168 h, 192 h, 216 h).
• Columns M–V: fresh weights (mg) of 10 dsegfp control larvae.
• Columns W–AF: fresh weights (mg) of 10 dsTai-injected larvae.

• Rows 16–27: “Multiple t test” summary (Time, Significant?, P value, means, mean difference, SE, t ratio, df).

(3) Figure 4C–4D – Survival and phenotypes

• Similar structure to sheet “Fig2” and “Fig3”:

• A table with “Days”, “dsegfp”, “dsTai”, and phenotype columns indicating
  numbers of individuals at different developmental outcomes used
  to draw Figure 4C and 4D.

Sheet “Fig5” – JH and 20E pathway genes after Tai RNAi (Figure 5)

This sheet contains qRT-PCR data for multiple genes, each with the same structure:
treatment vs. control with three biological replicates of 2^-ΔΔCt values.

(1) Figure 5A–5D – JH signaling genes

• Data block: rows 4–6, columns F–H, M–O, T–V, AB–AD.
• Row 4 (headers):

• F4: “HvmMet”
• G4–H4: “2-△△ct”
• M4: “Hvmkr-h1”
• N4–O4: “2-△△ct”
• T4: “HvmHairy”
• U4–V4: “2-△△ct”
• AB4: “HvmGroucho”
• AC4–AD4: “2-△△ct”

• Rows 5–6:

• For each gene:

• First column: treatment (“dsegfp”, “dsTai”).
• Next three columns: three biological 2^-ΔΔCt replicates.

(2) Figure 5E–5H – 20E signaling genes

• Data block: rows 21–23, columns F–H, M–O, T–V, AB–AD, etc.
• Row 21 (headers, example):

• F21: “HvmEcR”
• G21–H21: “2-△△ct”
• M21: “HvmUSP”
• N21–O21: “2-△△ct”
• T21: “HvmBrC”
• U21–V21: “2-△△ct”
• AB21: “HvmE74”
• AC21–AD21: “2-△△ct”
• Further genes (HvmE75, HvmE93, HvmHR4, HvmFtz-f1) are organized in the same way.

• Rows 22–23:

• For each gene:

• First column in the block: treatment (“dsegfp”, “dsTai”).
• Next three columns: three 2^-ΔΔCt replicate values.

(3) Additional rows (e.g., “Paired t test”, “P value”, “P value summary”)

• Rows 9–14 and 26–31 contain GraphPad Prism outputs for statistical comparisons
  between dsegfp and dsTai for each gene (P values, significance, t, and df).
• These are summary statistics, not raw qRT-PCR measurements.

1. Variable definitions (summary)

Across all sheets, the variables can be summarized as follows:

• SampleID / Stage / Tissue / Time:
  • Text labels identifying developmental stage, tissue, treatment, or time point
    (e.g., “L1D0”, “Head”, “0h”, “Day 5”, “dsegfp”, “dsTai”).
• 2-△Ct (three replicate columns):
  • Unitless relative expression values for HvTai (Figure 1).
  • Three biological replicates per sample.
• 2-△△Ct (three replicate columns):
  • Unitless relative expression values for each target gene (HvTai, HvMet,
    HvKr-h1, HvHairy, HvGroucho, HvEcR, HvUSP, HvBrC, HvE74, HvE75, HvE93,
    HvHR4, HvFtz-f1) normalized to reference genes and to a control group.
  • Three biological replicates per treatment.
• Fresh weight (mg):
  • Body weight of individual larvae at a given time point after dsRNA injection.
  • For each time point, 10 individuals per treatment group (dsegfp and dsTai).
• Survival and phenotype counts:
  • Number (or proportion) of larvae alive on each day after injection in each treatment group.
  • Number of individuals in different developmental categories
    (e.g., normal pupa, deformed larva, deformed pupa, prepupa), used to calculate
    survival curves and phenotype ratios.