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, Immunoprecipitation (IP) of chromatin-bound elongating RNAPII (RPB1 CTD-pS2) following treatment with UVC or formaldehyde (FA) at the indicated doses, representative of three independent experiments. b, Dsk2 pulldown of ubiquitylated proteins from RPE1 cells synchronized in G1 by serum starvation and treated with UVC or FA, or from cells released from a thymidine block into S-phase in the presence of deoxycytidine (dC) or 5-aza-dC, representative of three independent experiments. c, Dsk2 pulldown in WT and CSB−/− RPE1 cells synchronized in G1 by serum starvation and treated with 250 µM FA for the indicated times, representative of four independent experiments. d, Dsk2 pulldown in U2OS GFP–DNMT1 cells released from a single thymidine block into dC or 5-aza-dC for the indicated times, representative of three independent experiments. e,f, RPB1 degradation in cycloheximide-treated RPE1 cells following formaldehyde (e) or UVC (f) treatment in the presence or absence of MLN4924, an inhibitor of Cullin-dependent ubiquitylation, representative of three independent experiments. g,h, RPB1 degradation in cycloheximide-treated WT and CSB−/− (g) or CSA−/− (h) RPE1 cells at the indicated timepoints after a pulsed FA treatment. For g and h, GAPDH blot images are also shown alongside blots in Extended Data Fig. 3j,k, respectively, due to detection of RPB1 CTD-pS2 and RPB1 CTD-pS5 from the same experiment. i,j, Quantification of g and h, respectively; error bars ± s.e.m., n = 3 replicates. In b–d, *denotes polyubiquitylated RPB1 CTD-pS2. Source numerical data and unprocessed blots are available in Source data.
