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Fig. 6 | Genome Biology

Fig. 6

From: DNA Topoisomerase I differentially modulates R-loops across the human genome

Fig. 6

Top1 depletion triggers G0/G1 block and global replication timing delay. a Western blots showing the cellular response to Top1 depletion and camptothecin treatment with respect to γH2AX and markers of DNA damage signaling. b Total γH2AX immunofluorescence signal for control and Top1-depleted cells (n > 100 cells). c Cell cycle analysis for Top1-depleted cells and controls. Results are average of four experiments presented with standard deviation. d Representative images of ki-67 staining for Top1-depleted and control cells. Cells were counter-stained with DAPI. e Quantification of ki-67 staining (160 cells for each sample per experiment; two independent replicates). f Analysis of replication timing at a range of RLL loci in Top1-depleted cells and controls. The % of cells undergoing replication in each phase of the cell cycle was measured by the relative recovery of BrdU-labeled immunoprecipitated DNA across G1, early S (ES), late S (LS) and G2 phases. Error bars are SE of two replicates. Red and grey shading indicate genes with significant and non-significant replication timing delays, respectively. g Replication timing analysis of mitochondrial DNA

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