Fig. 5

Inhibition of protein synthesis affects CHK2 S516 phosphorylation levels. a G1-synchronized RPE-1 cells were left untreated (UT) or treated with a dose of 40 ng/ml of neocarzinostatin (NCS40) for 30 min; around 16 h after DNA damage, cycloheximide (CHX) was added at the indicated times before protein harvest. A representative western blot is shown (upper panel) together with the quantifications of four independent experiments (lower panel). b RPE-1 cells were treated as in a, but instead of using CHX, the MG132 was added at the indicated times before protein harvest. A representative western blot is shown (upper panel) together with the quantifications of three independent experiments (lower panel). In both quantifications, pCHK2 S516 levels where relativized with total CHK2 (blue lines), while cyclin D1 levels were relativized with HSP90 (green lines); only NCS-treated conditions were quantified. Error bars indicate standard deviations