Figure 7
PEITC induces γ-H2AX foci, activates ATM/CHK2, G2/M- and S-phase arrest and apoptosis. SK-BR-3 and A549 cells treated with PEITC or DMSO for 3 days were stained with anti-γ-H2AX antibody. (a) Merged images show cells stained with anti-γ-H2AX antibody (green) and DAPI (blue). All scale bars represents a size of 20 μm. (b) Percentage of cells with γ-H2AX foci (≤10 or >10, as indicated). (c) SK-BR-3 and A549 cells were treated with PEITC or DMSO for 4 h. Western blotting was performed using anti-pATM S1981 and anti-pCHK2 Thr68 antibodies. Blots were stripped and reprobed with anti-ATM and anti-CHK2 antibodies. (d) SK-BR-3 or (e) A549 cells were treated with PEITC, 10 μM Nutlin-3 or both for 24 h and analyzed by flow cytometry. (f) SK-BR-3 and A549 cells were treated with 4μM PEITC, 10 μM Nutlin-3 or both for 24 h. Apoptosis was measured by Annexin-V staining using a BD LSRFORTESSA instrument
