Fig. 6
No ATF2-dependent effect on DDR in HCT116 p53-/- cells upon 5-FU treatment. a, b, c Co-IP performed with an anti-ATF2 antibody in HT29 cells (a) or with an anti-Chk1 antibody (b) or with an anti-p53 antibody (c) in HT29, B5, and F10 cells. The Co-IP or whole cell extract (Input) was subjected to SDS–PAGE followed by Western blotting; ctrl*: 48 h DMSO treated cells for control, GAPDH was used as loading control for input. Band intensities were quantified using ImageJ analysis software, and ratios were calculated against the corresponding ctrl* band intensity. d ATF2 silencing through RNAi-mediated ATF2 knockdown. Representative Western blotting for ATF2, γ-H2AX, H2AX, PARP, cleaved PARP and p-Chk1Ser317, Chk1, p-ATRThr1989, ATR after treatment with various doses of 5-FU for 48 h; ctrl*: 48 h DMSO treated cells for control, GAPDH was used as loading control. Band intensities were quantified using ImageJ analysis software, and ratios were calculated against the GAPDH band intensity. For cleaved PARP, given ratios were calculated as cleaved PARP versus noncleaved PARP (ImageJ). e Co-IP performed with an anti-ATF2 antibody in HCT116 p53-/- cells. The Co-IP or whole cell extract (Input) was subjected to SDS–PAGE followed by Western blotting; ctrl*: 48 h DMSO treated cells for control, GAPDH was used as loading control for input. Band intensities were quantified using ImageJ analysis software, and ratios were calculated against the corresponding ctrl* band intensity