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. 2019 Nov 29;21(1):e48460. doi: 10.15252/embr.201948460

Figure EV3. PHF6 knockout cells display a similar phenotype as cells depleted for PHF6.

Figure EV3

  • A, B
    RPE1 cells were depleted of PHF6 using four different siRNA oligonucleotides. Checkpoint recovery after 5 Gy was analyzed by staining for MPM2 and flow cytometry (A), or cells were lysed and analyzed using Western blotting with the indicated antibodies (B). Error bars represent the SEM of three independent experiments. Statistical significance was determined using a two‐tailed, unpaired t‐test (***P < 0.001, ****P < 0.0001).
  • C
    U2OS WT and three different PHF6 knockout clones were lysed and analyzed by Western blotting with the indicated antibodies.
  • D
    U2OS WT and PHF6 knockout cells were irradiated at 2 Gy and fixed for immunofluorescence at 0, 1, and 16 h. Samples were stained with the indicated antibodies. Scale bar is 10 μm.
  • E
    Quantification of number of 53BP1 IRIF per cell of (D) in which the error bars represent the SD of three independent experiments. Statistical significance was determined using a two‐tailed, unpaired t‐test (**P < 0.01, ****P < 0.0001).
  • F
    U2OS cells were depleted of PHF6 with three different siRNA oligonucleotides. Cells were fixed, stained with propidium iodide, and analyzed by flow cytometry. Cell cycle distribution was determined using Flowlogic software. Error bars represent the SD of three independent experiments.
  • G
    U2OS WT and two different PHF6 knockout cell lines were fixed, stained with propidium iodide, and analyzed by flow cytometry. Cell cycle distribution was determined using Flowlogic software. Error bars represent the SD of three independent experiments.