Extended Data Fig. 3: CSB initiates a pathway that supports transcription recovery following DPC induction.
From: Transcription-coupled repair of DNA–protein cross-links depends on CSA and CSB
(a) Constitutive expression of GFP-DNMT1 compared to GFP in U2OS cells; full length GFP-DNMT1 is denoted by an asterisk (*). (b) Representative images from Proximity ligation assays (PLA) between GFP and RPB1 CTD-pS2 in U2OS cells constitutively expressing GFP-DNMT1 following release from thymidine block into 10 µM dC or 5-aza-dC for 1 h; scale bars = 10 µm. (c) Quantification of (b); data presented as mean ± SD, n = 3 replicates. (d-e) Quantification of Fig. 3e, f; degradation of RPB1 in RPE1 cells treated with FA (d) or UVC (e) in the presence or absence of MLN4924; data presented as mean ± SEM, n = 3 replicates. (f-g) RPB1 degradation in cycloheximide-treated WT and CSB−/− (f) or CSA−/− (g) RPE1 cells at the indicated time points after a UVC treatment. (h-i) Quantification of (f-g), respectively; data presented as mean ± SEM, n = 3 replicates. (j-k) RPB1 degradation in cycloheximide-treated WT and CSB−/− (j) or CSA−/− (k) cells treated with formaldehyde for the indicated time points. For (j) and (k), GAPDH blot images are also shown alongside blots in Fig. 3g, h, respectively, due to detection of RPB1 CTD-pS2 and RPB1 CTD-pS5 from the same experiment. (l-m) Quantification of (j) and (k), respectively; data presented as mean ± SEM, n = 3 replicates. Source numerical data and unprocessed blots are available in source data.
