Fig. 5: HDAC2 is the major deacetylase of KAT7 upon DNA damage stimulation.
a HCT116 cells were treated with TSA (3 μM, 12 h) and NAM (5 mM, 12 h). Cells were harvested to detect the acetylation levels of KAT7 by immunoprecipitation. b HCT116 cells were transfected with Flag-KAT7 plasmid and then stimulated with 40 μM etoposide for 8 h. KAT7-associated proteins were identified by MS. n = 1 biological replicate. The table lists the selected proteins identified by MS. The full protein list is provided in Supplementary Data File 1. c Flag-tagged HDAC2 were transfected in dose-dependent manner into HCT116 cells to confirm the effect of HDAC2 on KAT7 acetylation by immunoprecipitation. d Detection of KAT7 acetylation levels in HDAC2 stable knockdown HCT116 cells by immunoprecipitation. e HCT116 cells were transfected with Myc-KAT7 or Flag-HDAC2, and then the Myc-KAT7 and Flag-HDAC2 proteins were purified for in vitro deacetylation assay. KAT7 acetylation was detected by immunoprecipitation. f–h Exogenous, semi-exogenous and endogenous Co-IP were performed to detect the interaction between KAT7 and HDAC2 in HCT116 cells. i Endogenous Co-IP was performed to detect the interaction between KAT7 and HDAC2 in response to etoposide (40 μM, 8 h) treatment in HCT116 cells. j, k GST-HDAC2 FL and His-KAT7 FL were purified, and then western blotting was performed to detect the direct binding of KAT7 and HDAC2 in vitro. The bands were stained with Coomassie brilliant blue staining. # indicates the specific bands. The relative grayscale value of the band in the first lane (control group) was set as 1, and the ratio of the grayscale value of the other bands to the grayscale value of the first band was set as the relative grayscale value of the band. The experiments in Fig. 5a, c-k were three independent biological replicates. Source data are provided as a Source Data file.
