Extended Data Fig. 3: Phosphorylation of KAP1 at S824 but not S473 exhibits presentation at enhancers.
From: An eRNA transcription checkpoint for diverse signal-dependent enhancer activation programs
a) Venn Diagram illustrating the overlapped peaks between KAP1 and pS473-KAP1, or pS824-KAP1. b) Pie chart shows the genome-wide distribution of pS473-KAP1 and pS824-KAP1 peaks in E2 treated MCF7 cells by CUT&Tag. c) Heatmaps profiling of KAP1, pS473-KAP1, and pS824-KAP1 signals at KAP1-enriched E2 induced genes (n = 480), the data were generated from E2 treated or non- treated MCF7 cells. d) Browser track showing CUT&Tag signals at TFF1e, FOXC1e, RETe, and PVT1e loci. e) Immunoblot shows pS824-KAP1 level before or after E2 treatment in MCF7 cells. f) Immunoblot shows pS824-KAP1 level after endogenous KAP1 immunoprecipitation assay with or without E2 treatment in MCF7 cells. g) Endogenous DNA-PKcs co-IP assay showing the interaction between DNA-PKcs with ERα, KAP1, and pS824-KAP1. h) Endogenous pS824-KAP1 co-IP assay showing the interaction between pS824-KAP1 and DNA-PKcs. The experiments were repeated three times with similar results. i) RT-qPCR results show that the transcription of RETe and PVT1e are impaired with DNA-PKcs inhibitor NU7441 rather than the ATM inhibitor KU55933. Data presented as mean values ± s.d. from three independent biological replicates. P values are from two-tailed t-test.
