Figure 3. Disruption of the IR-Induced G₂/M Checkpoint Enhances Inflammatory Signaling Activation in MCF10A Cells.

(A) MCF10A cells were irradiated or left untreated with or without the indicated treatments, followed by fixation at the indicated times. Cells with micronuclei were quantified. Mean values and SEM are plotted (n = 3).

(B) MCF10A cells irradiated or left untreated with or without indicated inhibitors were fixed and subjected to cell cycle analysis by flow cytometry. Mean values and SEM are plotted (n = 3).

(C and D) MCF10A cells irradiated with the indicated dose or left untreated in the presence of the indicated inhibitors were collected at the specified time point for western blot.

(E) MCF10A I-PpoI cells were left untreated (NIR), irradiated with 10 Gy (IR), or treated with 4-OHT and shield-1 for 5 h (I-PpoI), and then maintained in medium with or without ATR inhibitor for 3 days before collection for western blot analysis.

(F) MCF10A AsiSI cells were left untreated (NIR), irradiated with 10 Gy (IR), induced with 4-OHT and shield-1 for 5 h(AsiSI), or cultured in the presence of aphidicolin (Aph), and then maintained in medium with or without ATR inhibitor for 3 days before collection for western blot analysis. For Aph-treated cells, 2.5 μM aphidicolin was included in the medium until cell collection 3 days later.

(G) MCF10A cells left untreated or irradiated with 10 Gy were maintained for 3 days in the presence or absence of ATR inhibitor before collection for western blot analysis.