KDM5 histone demethylases repress immune response via suppression of STING
Fig 5
KDM5B and KDM5C bind to the promoter of STING and directly suppress STING expression.
(A) Western blot (left panel) and RT-qPCR (right panel) analyses of STING in MCF7 cells with knockout of the indicated genes after treatment with DMSO or 1 μM KDM5-C70 for 3 days. (B, C) Western blot analysis of H3K4me3 (upper panel) and RT-qPCR analysis of STING (lower panel) in MCF7 cells (panel B) or in BT474 cells (panel C) after treatment with the indicated compounds for 3 days. The concentration of OICR-9429 was 20 μM. (D) H3K4me3 ChIP-qPCR analysis at the promoter of STING, OAS2, or IFI44L in control or STING knockout MCF7 cells treated with DMSO or 1 μM KDM5-C70 for 1 day or 6 days. (E) KDM5A and KDM5B ChIP-qPCR analysis of MCF7 cells at the STING and NDUFA9 promoters, or downstream of the last STING exon as NC. (F) Analysis of ChIP-seq data for KDM5B binding at the STING genomic region in K562 cells (GSE29611, upper panel) and KDM5C in ZR-75-30 cells (GSE71327, lower panel) [42]. Representative data from triplicate experiments are shown. Error bar denotes SEM. #p < 0.01 for the comparisons shown in panel A–C, for inhibitors versus DMSO (panel D), and for KDM5A or KDM5B ChIP versus IgG ChIP (panel E). The numerical values used to generate graphs in panel A–E are available in S1 Data. ChIP, chromatin immunoprecipitation; ChIP-seq, chromatin immunoprecipitation sequencing; IgG, immunoglobulin G; NC, negative control; RT-qPCR, RT-qPCR, reverse transcription followed by quantitative PCR; STING, stimulator of interferon genes.