Fig. 4: lncREST regulates fork progression during replication stress.

a Experimental setup for replication fork analysis in lncREST KD cells, and representative fields of DNA fibers immunofluorescence. Scalebar: 20 μm. b Tract length frequency analysis in the IdU pulse (left) and CldU + HU (right). For each condition, Kernel density estimates were plotted. At least 250 fibers for each sample were analyzed. *p = 0.0122 in CTRL vs lncREST LNA1 CldU; *p = 0.0386 in CTRL vs lncREST LNA2 CldU (two-tailed unpaired t-test performed on average tract size for the different replicates; n = 3 per group). c Experimental setup for replication fork analysis of control or lncREST KD cells treated with S1 nuclease, and dot plot and median of CldU/IdU ratio from 40 to 200 fibers per replicate, n = 3. ***p < 0.001, ****p < 0.0001, by two-tailed Mann-Whitney test. d qRT-PCR showing lncREST expression in HCT116 wt and lncREST-KO1 cells treated with EV (pcDNA) or pcDNA-lncREST plasmid. **p < 0.01, ***p < 0.001, ****p < 0.0001 (mean ± SD, two-tailed unpaired t-test. n = 3 per group). e Experimental setting for replication fork analysis and representative image of fiber assay in HCT116 wt and lncREST-KO cells treated with EV (pcDNA) or pcDNA-lncREST plasmid (left), and dot blot with median of IdU (green) tracts (right); at least 150 fibers for each replicate were scored (n = 3) ****p < 0.0001, by two-tailed Mann-Whitney test. Source data are provided as a Source Data file.