Extended Data Fig. 4: INTS12 and its phosphorylation are required for efficient transcriptional recovery following UV irradiation.
From: Integrator subunit INTS12 links ribotoxic stress to transcription-coupled nucleotide excision repair
a, Clonogenic survival of control, INTS12 KO and CSB KO cells treated with indicated doses of 4-NQO. Graphs represent the mean ± s.d. from n = 3 repeated experiments and were normalized to the untreated colony number, which was set at 100%. b, c, Cell survival of control and INTS12 KO cells treated with indicated doses of IR (b) or Calicheamicin (c). Graphs represent the mean ± s.d. from n = 3 repeated experiments and were normalized to the untreated cell number, which was set at 100%. d, e, INTS12 KO cells reconstituted with either INTS12-WT or INTS12-3A were treated with indicated doses of UV (d) or 4-NQO (e) and subjected to clonogenic survival assays. Graphs represent the mean ± s.d. from n = 3 repeated experiments and were normalized to the untreated colony number, which was set at 100%. f, Quantification of fluorescence intensity per cell for data shown in Fig. 3b and are representative of three independent experiment (n = 238). Red lines indicate the mean intensity in each group. g, Transcription recovery in INTS12 KO cells reconstituted with either INTS12-WT or INTS12-3A following UV irradiation was measured by 5-EU labeling of newly synthesized RNA. Right: Quantification of fluorescence intensity per cell and are representative of three independent experiment (n = 216). Red lines indicate the mean intensity in each group. Scale bar, 50 μm. h, Transcription recovery in control, ZAK KO and ZAKi-treated (Nilotinib, 1 µM, pre-1h) cells following UV irradiation was measured by 5-EU labeling of newly synthesized RNA. Bottom: Quantification of fluorescence intensity per cell and are representative of three independent experiment (n = 242). Red lines indicate the mean intensity in each group. Scale bar, 20 μm. i, Metaplots showing the average TT-seq signal levels of genes between 10-25 kb, 25-50 kb, and 50-100 kb in control and INTS12 KO cells after mock or UV treatment, followed by recovery for the indicated times.
