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. 2019 Aug 19;38(20):e101443. doi: 10.15252/embj.2018101443

Figure EV2. Loss of cyclin F results in S phase accumulation of DNA damage (related to Figs 2 and 3).

Figure EV2

  1. U‐2‐OS cells transfected with the indicated siRNA and treated with Chk1i (LY2603618) for 20 h were harvested and lysed using SDS. Indicated proteins were resolved by SDS–PAGE and detected by Western blot (WB).
  2. Percentage (%) of γ‐H2AX‐positive cells in early S cells after Chk1i treatment (LY2603618). Early S phase cells were considered having 2n DAPI content and EdU+. Data from at least three independent experiments were plotted with mean % ± SD.
  3. Percentage (%) of γ‐H2AX‐positive cells in mid‐late S phase after Chk1i (LY2603618) treatment. Mid‐late S phase cells were considered having DAPI 4n staining and EdU+. Data from at least 3 independent experiments were plotted with mean % ± SD.
  4. Representative plots of HeLa and CCNF K/O cells treated with 1 μM Chk1i for the indicated hours (h). Half an hour before harvesting, cells were incorporated with EdU (10 μM as final concentration). EdU by click reaction, γ‐H2Ax and RPA by immunostaining, and DAPI staining were detected by FACS analysis.