Extended Data Fig. 6: Downstream NER factors are dispensable for transcription recovery after DPC induction.
From: Transcription-coupled repair of DNA–protein cross-links depends on CSA and CSB
(a) Representative images from RRS assays in WT, CSB−/− and UVSSA−/− RPE1 cells treated with formaldehyde; scale bars = 10 µm, images representative of 3 independent experiments. (b) Dsk2 pulldown in G1-synchronised WT, CSB−/− and UVSSA−/− RPE1 cells treated with formaldehyde for the indicated times; data representative of 3 independent experiments. (c) Formaldehyde-induced RPB1 degradation in cycloheximide-treated WT and UVSSA−/− cells released from FA treatment for the indicated times. (d) Quantification of (c); error bars ± SEM, n = 3 replicates. (e) Representative images from RRS assays in XPC−/−, XPC−/−/CSB−/− and XPC−/−/XPA−/− RPE1 cells treated with formaldehyde; scale bars = 10 µm. (f) Western blot analysis of candidate XPG knockout RPE1 clones. (g) Sanger sequencing result from genotyping of RPE1 XPG−/− clone 21 used following PCR amplification of the targeted gDNA sequence. (h-i) Alamar blue assays in WT, CSB−/−, ERCC1−/− and XPG−/− RPE1 cells treated with formaldehyde (e) or 5-aza-dC (f); data presented as mean ± SD, n = 3 replicates. Source numerical data and unprocessed blots are available in source data.
