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KDM5 histone demethylases repress immune response via suppression of STING

Fig 1

Inhibition of KDM5 demethylases activates interferon-induced genes.

(A) Western blot analysis of histone modifications in MCF7 cells treated with 1 μM KDM5-C70, 10 μM Dong-A-167, 10 μM GDC-50, or 10 μM CPI-48 for 3 days. (B) GSEA of RNA-seq data from MCF7 cells treated with 3 μM KDM5-C70 or CPI-48 for 6 days. (C, D) RT-qPCR (panel C) and western blot (panel D) analyses of MCF7 cells treated with 1 μM KDM5-C70, 10 μM Dong-A-167, 10 μM GDC-50, or 10 μM CPI-48 for 6 days. (E, F) Western blot (panel E) and RT-qPCR (panel F) analyses of MCF7 cells with stable knockout of the indicated genes using the CRISPR/Cas9 system. Control 1, empty vector; control 2, scrambled sequence. KDM5BC, 2 sgRNAs targeting KDM5B and KDM5C. (G) Western blot analysis of MCF7 cells after transfection with indicated plasmids. Representative data from triplicate experiments are shown. Error bar denotes SEM. #p < 0.01 for inhibitors versus DMSO (panel C), and for knockout sgRNA versus average of 2 control sgRNAs (panel F). The numerical values used to generate graphs in panel C and F are available in S1 Data. CRSPR/Cas9, clustered regular interspaced short palindromic repeats/CRISPR-associated protein 9; FDR q, false discovery rate q value; GSEA, gene set enrichment analysis; NES, normalized enrichment score; NOM p, nominal p value; NS, nonspecific band; RNA-seq, RNA sequencing; RT-qPCR, reverse transcription followed by quantitative PCR; sgRNA, single guide RNA.

Fig 1

doi: https://doi.org/10.1371/journal.pbio.2006134.g001