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. 2019 Sep 2;38(20):e101430. doi: 10.15252/embj.2018101430

Figure EV3. Failure to degrade E2F7/8 caused cell cycle delay and DNA damage accumulation.

Figure EV3

  1. Confirmation of efficient deletion of E2F7 and E2F8 in RPE‐FUCCI cells utilizing CRISPR–CAS9 technology. Cells were transduced with Flag‐tagged Cas9, and either a construct lacking sgRNA (Ctrl), or a construct containing sgRNA directed against E2F7 and E2F8 (7/8 KO). Protein levels were analyzed by immunoblotting.
  2. Depletion of cyclin F stalls the cell cycle at late S/G2. Each bar in the histogram shows the time that each individual of cell has spent from G1/S to Mitosis (correspond to Fig 4F). Red (G1), Green (S/G2) and Gray (M/G0) indicate the cell cycle stage of each cell after 24 h live cell imaging.
  3. Over expression of E2F7R894A mutant caused DNA damage. HeLa/TO cells were arrested with HU for 16 h and then released into fresh medium containing doxycycline. Protein samples were harvested every 3 h, and γ‐H2AX was measured by immunoblotting (left). Quantifications were performed based on two independent experiments (right). Bars and error bars represent mean ± SEM.

Source data are available online for this figure.