Fig. 6

FOXD1 is necessary for maintaining DNA integrity following mitosis. a Representative TUNEL (red) staining of 786-O versus 786-O^FOXD1null, counterstained with DAPI (blue) . Nuclear TUNEL foci were counted and compared using Student's t-test (N = 3). *p < 0.05 b Representative γH2AX (red) immunocytochemistry of 786-O versus 786-O^FOXD1null cells, counterstained with DAPI (blue). Nuclear γH2AX foci were counted and compared using Student’s t-test (N = 3). *p < 0.05 c EdU pulse-chase cell cycle progression analysis. Cells in G2 were labeled with EdU following 8 h incubation with EdU. Labeled cells were analyzed for pH3 expression every 3 h post-labeling using immunocytochemistry. One hundred EdU+ cells were analyzed (N = 3). d Analysis of EdU labeled cells in G2 for γH2AX expression every 2 h post- labeling by immunocytochemistry. One hundred EdU+ cells were analyzed (N = 3). e Immunocytochemistry analysis of γH2AX (green) at different phases of the cell cycle, costained with DAPI (gray) and phalloidin (blue). Early M represented by cells in metaphase and late M representing cells undergoing cytokinesis and entering post-mitosis G1. f Correlation of mean FOXD1 expression against all genes found in the KIRC TCGA database. Ranked values scored using the Pearson correlation coefficient. g Gene set enrichment analysis of ranked FOXD1-correlated genes shows enrichment of REACTOME pathways related to mitotic entry and progression