Extended Data Fig. 3: INTS12 contributes to Integrator recruitment to CSB.
From: Integrator subunit INTS12 links ribotoxic stress to transcription-coupled nucleotide excision repair
a, Venn diagram showing the number of proteins identified by MS from the INTS12-WT or 3 A interactome. b, Gene Ontology analysis of the MS-identified proteins that specifically interacted with INTS12-WT after UV irradiation. c, INTS12 KO cells reconstituted with stably expressed WT or 3 A INTS12-Flag were untreated or treated with 4-NQO. Chromatin fractions (Input) and anti-Flag immunoprecipitates (IPs) were analyzed by WB. d, Purified recombinant CSB and INTS12 proteins were analyzed by SDS-PAGE and visualized by Coomassie blue staining. e, Control or CSB KO cells expressing INTS12-Flag were either untreated or treated with UV as indicated and then analyzed as in c. f, g, Control or INTS12 KO cells expressing Flag-PAF1 or not (-) were untreated (g) or treated with UV as indicated (f) and then analyzed as in c. h–j, Control or INTS12 KO cells expressing INTS3-Flag (h), INTS5-Flag (i), INTS10-Flag (j) or not (-) were untreated or treated with UV as indicated and then analyzed as in c. k, l, Control or INTS12 KO expressing INTS5-Flag (k) or not (l) were treated with DMSO or 4-NQO (5 µM, 1 h) and subject to PLA using the indicated antibodies. Right: quantification of PLA foci per nucleus (n = 141). P < 0.0001. m, HeLa cells expressing INTS10-Flag together with either INTS3-Strep or INTS3 (∆CMBM)-Strep were lysed to prepare for the whole cell extracts (WCEs). WCEs (Input) and anti-strep immunoprecipitates (IPs) were analyzed by WB. n, HeLa cells expressing INTS3-Strep or INTS3 (∆CMBM)-Strep were either untreated or treated with UV as indicated and then analyzed as in c. For k and l, box plots show the minimum, quartiles and maximum. Scale bar, 10 μm. Statistical analysis was performed using two-tailed unpaired t-tests. All Western blots are representative of three independent experiments.
