Dynamic post-translational modification profiling of M. tuberculosis-infected primary macrophages
Data files
Jan 30, 2020 version files 157.01 MB
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Figure_4-Figure_S1_Source_Data_1_Panel_A_anti-FLAG.TIF
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Figure_4-Figure_S1_Source_Data_2_Panel_A_anti-actin.tif
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Figure_4-Figure_S1_Source_Data_3_Panel_B_anti-FLAG.TIF
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Figure_4-Figure_S1_Source_Data_4_Panel_B_anti-actin.tif
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Figure_4-Source_Data_1_control.tif
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Figure_4-Source_Data_2_p62.tif
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Figure_4-Source_Data_3_optineurin.tif
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Figure_4-Source_Data_4_Tax1bp1.tif
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Figure_4-Source_Data_5_bnip3.tif
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Figure_4-Source_Data_6_bnip3l.tif
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Figure_4-Source_Data_7_fundc1.tif
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Figure_4-Source_Data_8_nbr.tif
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Figure_4-Source_Data_9.csv
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Figure_5-Fig_S1_Source_Data_28.csv
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Figure_5-Figure_S1_Source_Data_1_Scramble_anti-Ub_488.tiff
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Figure_5-Figure_S1_Source_Data_10_Scramble_anti-pTBK1_DAPI.tiff
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Figure_5-Figure_S1_Source_Data_11_Scramble_anti-pTBK1_488.tiff
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Figure_5-Figure_S1_Source_Data_12_Scramble_anti-pTBK1_mCherry.tiff
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Figure_5-Figure_S1_Source_Data_13_Tax1bp1_exon_10_anti-pTBK1_DAPI.tiff
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Figure_5-Figure_S1_Source_Data_14_Tax1bp1_exon_10_anti-pTBK1_488.tiff
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Figure_5-Figure_S1_Source_Data_15_Tax1bp1_exon_10_anti-pTBK1_mCherry.tiff
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Figure_5-Figure_S1_Source_Data_16_Tax1bp1_exon_9_anti-pTBK1_488.tiff
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Figure_5-Figure_S1_Source_Data_17_Tax1bp1_exon_9_anti-pTBK1_DAPI.tiff
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Figure_5-Figure_S1_Source_Data_18_Tax1bp1_exon_9_anti-pTBK1_mCherry.tiff
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Figure_5-Figure_S1_Source_Data_19_Scramble_anti-p62_DAPI.tiff
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Figure_5-Figure_S1_Source_Data_2_Scramble_anti-Ub_DAPI.tiff
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Figure_5-Figure_S1_Source_Data_20_Scramble_anti-p62_488.tiff
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Figure_5-Figure_S1_Source_Data_21_Scramble_anti-p62_mCherry.tiff
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Figure_5-Figure_S1_Source_Data_22_Tax1bp1_exon_10_anti-p62_DAPI.tiff
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Figure_5-Figure_S1_Source_Data_23_Tax1bp1_exon_10_anti-p62_488.tiff
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Figure_5-Figure_S1_Source_Data_24_Tax1bp1_exon_10_anti-p62_mCherry.tiff
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Figure_5-Figure_S1_Source_Data_25_Tax1bp1_exon_9_anti-p62_DAPI.tiff
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Figure_5-Figure_S1_Source_Data_26_Tax1bp1_exon_9_anti-p62_488.tiff
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Figure_5-Figure_S1_Source_Data_27_Tax1bp1_exon_9_anti-p62_mCherry.tiff
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Figure_5-Figure_S1_Source_Data_3_Scramble_anti-Ub_mCherry.tiff
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Figure_5-Figure_S1_Source_Data_4_Tax1bp1_exon_10_anti-Ub_DAPI.tiff
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Figure_5-Figure_S1_Source_Data_5_Tax1bp1_exon_10_anti-Ub_488.tiff
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Figure_5-Figure_S1_Source_Data_6_Tax1bp1_exon_10_anti-Ub_mCherry.tiff
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Figure_5-Figure_S1_Source_Data_7_Tax1bp1_exon_9_anti-Ub_mCherry.tiff
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Figure_5-Figure_S1_Source_Data_8_Tax1bp1_exon_9_anti-Ub_DAPI.tiff
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Figure_5-Figure_S1_Source_Data_9_Tax1bp1_exon_9_anti-Ub_488.tiff
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Figure_5-Source_Data_1_WT_anti-Ub_DAPI.tiff
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Figure_5-Source_Data_10_WT_anti-pTBK1_DAPI.tiff
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Figure_5-Source_Data_11_WT_anti-pTBK1_488.tiff
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Figure_5-Source_Data_12_WT_anti-pTBK1_mCherry.tiff
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Figure_5-Source_Data_13_p62_mutant_anti-pTBK1_DAPI.tiff
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Figure_5-Source_Data_14_p62_mutant_anti-pTBK1_488.tiff
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Figure_5-Source_Data_15_p62_mutant_anti-pTBK1_mCherry.tiff
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Figure_5-Source_Data_16_Tax1bp1_mutant_anti-pTBK1_DAPI.tiff
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Figure_5-Source_Data_17_Tax1bp1_mutant_anti-pTBK1_488.tiff
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Figure_5-Source_Data_18_Tax1bp1_mutant_anti-pTBK1_mCherry.tiff
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Figure_5-Source_Data_19_WT_anti-p62_DAPI.tiff
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Figure_5-Source_Data_2_WT_anti-Ub_488.tiff
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Figure_5-Source_Data_20_WT_anti-p62_488.tiff
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Figure_5-Source_Data_21_WT_anti-p62_mCherry.tiff
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Figure_5-Source_Data_22_p62_mutant_anti-p62_DAPI.tiff
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Figure_5-Source_Data_23_p62_mutant_anti-p62_488.tiff
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Figure_5-Source_Data_24_p62_mutant_anti-p62_mCherry.tiff
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Figure_5-Source_Data_25_Tax1bp1_mutant_anti-p62_DAPI.tiff
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Figure_5-Source_Data_26_Tax1bp1_mutant_anti-p62_488.tiff
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Figure_5-Source_Data_27_Tax1bp1_mutant_anti-p62_mCherry.tiff
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Figure_5-Source_Data_28.csv
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Figure_5-Source_Data_3_WT_anti-Ub_mCherry.tiff
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Figure_5-Source_Data_4_p62_mutant_anti-Ub_DAPI.tiff
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Figure_5-Source_Data_5_p62_mutant_anti-Ub_488.tiff
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Figure_5-Source_Data_6_p62_mutant_anti-Ub_mCherry.tiff
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Figure_5-Source_Data_7_Tax1bp1_mutant_anti-Ub_DAPI.tiff
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Figure_5-Source_Data_8_Tax1bp1_mutant_anti-Ub_488.tiff
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Figure_5-Source_Data_9_Tax1bp1_mutant_anti-Ub_mCherry.tiff
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Figure_6-Figure_S1_Source_Data_1_WT_unstimulated_Alexa_6475.tiff
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Figure_6-Figure_S1_Source_Data_10_p62_unstimulated_Alexa_6475.tiff
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Figure_6-Figure_S1_Source_Data_11_p62_unstimulated_DAPI5.tiff
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Figure_6-Figure_S1_Source_Data_12_p62_unstimulated_mCherry.tiff
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Figure_6-Figure_S1_Source_Data_13_p62_IFNg_post_Alexa_6476.tiff
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Figure_6-Figure_S1_Source_Data_14_p62_IFNg_post_DAPI6.tiff
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Figure_6-Figure_S1_Source_Data_15_p62_IFNg_post_mCherry6.tiff
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Figure_6-Figure_S1_Source_Data_16_p62_IFNg_pre_post_Alexa_6472.tiff
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Figure_6-Figure_S1_Source_Data_17_p62_IFNg_pre_post_DAPI2.tiff
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Figure_6-Figure_S1_Source_Data_18_p62_IFNg_pre_post_mCherry2.tiff
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Figure_6-Figure_S1_Source_Data_19_Tax1bp1_unstimulated_Alexa_6474.tiff
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Figure_6-Figure_S1_Source_Data_2_WT_unstimulated_mCherry5.tiff
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Figure_6-Figure_S1_Source_Data_20_Tax1bp1_unstimulated_DAPI4.tiff
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Figure_6-Figure_S1_Source_Data_21_Tax1bp1_unstimulated_mCherry4.tiff
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Figure_6-Figure_S1_Source_Data_22_Tax1bp1_IFNg_post_Alexa_6475.tiff
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Figure_6-Figure_S1_Source_Data_23_Tax1bp1_IFNg_post_DAPI5.tiff
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Figure_6-Figure_S1_Source_Data_24_Tax1bp1_IFNg_post_mCherry5.tiff
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Figure_6-Figure_S1_Source_Data_25_Tax1bp1_IFNg_pre_post_Alexa_6471.tiff
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Figure_6-Figure_S1_Source_Data_26_Tax1bp1_IFNg_pre_post_DAPI1.tiff
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Figure_6-Figure_S1_Source_Data_27_Tax1bp1_IFNg_pre_post_mCherry1.tiff
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Figure_6-Figure_S1_Source_Data_28.csv
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Figure_6-Figure_S1_Source_Data_3_WT_unstimulated_DAPI.tiff
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Figure_6-Figure_S1_Source_Data_4_WT_IFNg_post_Alexa_6473.tiff
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Figure_6-Figure_S1_Source_Data_5_WT_IFNg_post_mCherry3.tiff
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Figure_6-Figure_S1_Source_Data_6_WT_IFNg_post_DAPI3.tiff
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Figure_6-Figure_S1_Source_Data_7_WT_IFNg_pre-_post-_Alexa_6473.tiff
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Figure_6-Figure_S1_Source_Data_8_WT_IFNg_pre-_post-_DAPI3.tiff
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Figure_6-Figure_S1_Source_Data_9_WT_IFNg_pre-_post-_mCherry3.tiff
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Figure_6-Figure_S2_Source_Data_1_Griess.csv
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Figure_6-Figure_S2_Source_Data_2_Griess_(2).csv
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Figure_6-Figure_S2_Source_Data_3_ISRE.csv
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Figure_6-Figure_S2_Source_Data_4_IL-12.csv
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Figure_6-Figure_S2_Source_Data_5_IL-6.csv
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Figure_6-Figure_S2_Source_Data_6_TNFa.csv
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Figure_6-Source_Data_1_WT_mCherry2.tiff
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Figure_6-Source_Data_10_Scramble_Alexa_647.tiff
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Figure_6-Source_Data_11_Scramble_DAPI.tiff
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Figure_6-Source_Data_12_Scramble_mCherry.tiff
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Figure_6-Source_Data_13_Tax1bp1_Exon_10_DAPI.tiff
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Figure_6-Source_Data_14_Tax1bp1_Exon_10_mCherry.tiff
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Figure_6-Source_Data_15_Tax1bp1_Exon_10_Alexa_647.tiff
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Figure_6-Source_Data_16_Tax1bp1_Exon_9_DAPI.tiff
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Figure_6-Source_Data_17_Tax1bp1_Exon_9_mCherry.tiff
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Figure_6-Source_Data_18_Tax1bp1_Exon_9_Alexa_647.tiff
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Figure_6-Source_Data_19_Colocalization.csv
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Figure_6-Source_Data_2_WT_DAPI2.tiff
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Figure_6-Source_Data_20.csv
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Figure_6-Source_Data_21.csv
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Figure_6-Source_Data_22.csv
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Figure_6-Source_Data_3_WT_Alexa_6472.tiff
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Figure_6-Source_Data_4_p62_Alexa_647.tiff
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Figure_6-Source_Data_5_p62_DAPI.tiff
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Figure_6-Source_Data_6_p62_mCherry.tiff
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Figure_6-Source_Data_7_Tax1bp1_Alexa_647.tiff7.tiff
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Figure_6-Source_Data_8_Tax1bp1_mCherry.tiff7.tiff
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Figure_6-Source_Data_9_Tax1bp1_DAPI.tiff7.tiff
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Abstract
Macrophages are highly plastic cells with critical roles in immunity, cancer, and tissue homeostasis, but how these distinct cellular fates are triggered by environmental cues is poorly understood. To uncover how primary murine macrophages respond to bacterial pathogens, we globally assessed changes in post-translational modifications of proteins during infection with Mycobacterium tuberculosis, a notorious intracellular pathogen. We identified hundreds of dynamically regulated phosphorylation and ubiquitylation sites, indicating that dramatic remodeling of multiple host pathways, both expected and unexpected, occurred during infection. Most of these cellular changes were not captured by mRNA profiling, and included activation of ubiquitin-mediated autophagy, an evolutionarily ancient cellular antimicrobial system. This analysis also revealed that a particular autophagy receptor, TAX1BP1, mediates clearance of ubiquitylated Mtb and targets bacteria to LC3-positive phagophores. These studies provide a new resource for understanding how macrophages shape their proteome to meet the challenge of infection.
Immunofluorescence microscopy images were obtained with the Opera Phenix (Perkin Elmer) at 63X magnification.
Immunoblot images were obtained with a Li-Cor Odyssey instrument.
Please see the manuscript file for details about staining protocols.