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. 2018 Jan 23;37(5):e97877. doi: 10.15252/embj.201797877

Figure EV3. Prevention of Chk1‐mediated phosphorylation of atypical E2Fs delays cell cycle progression and reduces cell survival.

Figure EV3

  1. Cell cycle profile analysis by flow cytometry confirmed that over‐expression of non‐phosphorylatable forms of E2F7 or E2F8 delays cell cycle progression after HU release. HeLa/TO cells expressing wild‐type or mutant forms of E2F7/8 were harvested at 0, 9, and 14 h after HU release and doxycycline induction.
  2. Cell cycle profile analysis by flow cytometry confirmed that over‐expression of non‐phosphorylatable forms of E2F7 or E2F8 delays cell cycle progression after double thymidine block/release. HeLa/TO cells expressing wild‐type or mutant forms of E2F7/8 were harvested at 0, 9, and 14 h after double thymidine block/release and doxycycline induction.
  3. Representative montage figures from the live cell imaging show that over‐expression of alanine forms of E2F7/8 induces cell blebbing (arrows), interpreted as cell death. Scale bar: 10 μm.
  4. FACS sorting of cells expressing wild‐type and mutant E2F7 and E2F8. Sorting gates (rectangles) were set to isolate cells with comparable levels of expression from all E2F7/8‐EGFP‐expressing (Dox) cell lines.