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. 2020 Jun 2;39(14):e104036. doi: 10.15252/embj.2019104036

Figure 2. G2/M DNA damage checkpoint abrogation is required for interferon signaling induced by ATR inhibition + ionizing radiation (IR).

Figure 2

  1. MCF10A WT cells were pretreated with AZD6738 (250 nM) and RO‐3306 (9 μM) for 2 h, and then irradiated with 20 Gy; 3 days later, cells were harvested for immunoblotting with indicated antibodies.
  2. Same condition as described in A, except that RO‐3306 is replaced by PD‐0332991 (1 μM).
  3. MCF10A WT cells were pretreated with DMSO or AZD6738 (250 nM) for 2 h, and then irradiated with 20 Gy; 24 h later, cell cycle distribution was measured by flow cytometry. EdU (10 μM) was added into medium 2 h before harvesting for flow cytometry.
  4. MCF10A WT cells were pretreated with DMSO or AZD6738 (250 nM) for 2 h, and then irradiated with 20 Gy, and cells were harvested at indicated time points for immunoblotting.
  5. MCF10A control cells and CDC25C‐WT/constitutively active mutant CDC25C‐S216A overexpressed cells were pretreated with DMSO or AZD6738 (250 nM) for 2 h, and then irradiated with 20 Gy; 24 h later, cells were harvested for immunoblotting with indicated antibodies.
  6. Same condition as indicated in E, except that cells were harvested 72 h later.