Fig. 4: MCM on chromatin after POLE1 depletion.
From: The non-catalytic role of DNA polymerase epsilon in replication initiation in human cells
a, b Clone 16 cells were treated for 16 h with DMSO or dox/aux, followed chromatin extraction and MCM7 immunostaining. Flow cytometry plots of DNA/MCM7 staining (a) or MCM7 histograms (b) are shown. c–g Clone 16 cells treated for 16 h with DMSO or dox/aux were pulse labeled with thymidine analog EdU for 15 min prior to processing for super-resolution imaging. Representative multicolor super-resolution images (c) of PCNA (blue), MCM (green), and extent of EdU (red) incorporation in S-phase nuclei. Scale bar, 1500 nm. Quantitation of EdU (d) and MCM (e) detected per focus normalized to DMSO-treated clone 16 control, based on at least 3 independent experiments (For EdU per focus, n = 131, 109, and for MCM per focus n = 95, 83 for DMSO and dox/aux treated clone 16 cells. Student’s t-test was used for analyses, mean + SD and the p values are shown. Quantitation of PCNA detected per focus (f) and PCNA clusters (g), normalized to DMSO-treated clone 16 cells based on at least 2 independent experiments. (PCNA per focus, n = 126, 106; PCNA cluster density, n = 139, 120; student’s t-test was used for analyses, mean + SD and p values are shown). Source data are provided as a Source data file.
