Fig. 5: Loss of OFD1 in transformed cancer cells leads to cytokinesis failure and mitotic cell death. | Nature Communications

Fig. 5: Loss of OFD1 in transformed cancer cells leads to cytokinesis failure and mitotic cell death.

From: An actin filament branching surveillance system regulates cell cycle progression, cytokinesis and primary ciliogenesis

Fig. 5

a Proliferation curves of RPE1 cells expressing SV40 T Antigen with or without knockdown of OFD1. Data shown represented as mean values ± SD, error bar was defined as SD. b Proliferation curves of CEP164−/− RPE1/TAg cells with or without knockdown of OFD1. 300 cells examined over three independent experiments. Data shown represented as mean values ± SD, error bar was defined as SD. c Flow cytometry analysis of the DNA content of RPE1/TAg cells treated as indicated. Cells were stained with PI to show DNA content. d Mitotic events of WT (Left Panel) and OFD1-depleted (Right Panel) RPE1/TAg cells were monitored by phase-contrast time-lapse microscopy. Yellow arrows and arrowheads point at control cells undergoing normal M-phase. White arrows indicate OFD1-depleted cells undergoing mitotic cell death. Orange and red arrows point at OFD1-depleted cells with mitotic failure. Nuclei were labeled with Hoechst. e Quantitation of the duration of M phase in cells transfected with indicated siRNAs. 50 cells examined over three independent experiments, two-tailed unpaired student’s t-test. f Mitotic events of WT (Upper Panel) and OFD1-depleted (Lower Panel) RPE1/Tag cells were monitored by fluorescence time-lapse microscopy. RPF-LifeAct (red) and GFP-tubulin (green) were used to visualize F-actin and microtubules, respectively. Arrowheads point at the position of actomyosin rings. g Quantitation of the ratio of fluorescence signal of F-actin at the equator and pole in cells transfected with indicated siRNAs. Cells (n ≥ 50) examined over three independent experiments, data shown represented as mean values ± SD, error bar was defined as SD, two-tailed unpaired student’s t-test. h Quantitation of the indicated mitotic events of cells transfected with indicated siRNAs. Data shown represent mean ± SD. 300 cells examined over three independent experiments, data shown represented as mean values ± SD, error bar was defined as SD, two-tailed unpaired student’s t-test.

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