Figure 2

Checkpoint activation in response to NEU-induced damage is independent of mismatch repair. (A) siLacZ, siMsh2 and siMsh6 transfected HeLa cells were treated with 10 mM NEU for 2 hours and lysates were analysed for activation of checkpoint proteins by immunoblotting. (B) HCT116, a MLH1-deficient cell line, was treated with 0, 2, 6 and 10 mM NEU for 2 hours and lysates were analysed for activation of checkpoint proteins by immunoblotting. (C) DLD1, a MSH6 deficient cell line, was treated with 0, 2, 6 and 10 mM NEU for 2 hours and lysates were analysed for activation of checkpoint proteins by immunoblotting. (D) DLD1 cells were treated with 10 mM NEU in presence of 10 μM KU 55933 (ATM inhibitor) for 2 hours and lysates were analysed for activation of checkpoint proteins by immunoblotting. (E) DLD1 cells were treated with 10 mM NEU for 1 hour, fixed and analysed for formation of γH2AX foci by immunostaining. DMSO was used as negative control and 200 ng/ml NCS, an IR-mimetic drug, was used as positive control. Scale bar: 20 μM.