Figure 2. p400 depletion leads to ROS accumulation and DNA damage induction.

(A) U2OS cells were transfected using the indicated siRNA. 48 and 72 h following transfection (as indicated), cells were collected and ROS levels were measured by flow cytometry. The mean and standard deviation (SD) from 3 independent experiments are shown (after standardisation relative to 1 for cells transfected with the control siRNA). (B) U2OS cells were transfected using the indicated siRNA and 48 h following transfection, cells were collected and ROS levels were measured by flow cytometry. The mean and SD from 3 independent experiments are shown (after standardisation relative to 1 for cells transfected with the control siRNA). (C) MEFs derived from heterozygous embryos in which one p400 allele was targeted (p400Mut/+)[19] or from control wild type embryos (wt) were treated or not, as indicated, with 10 mM of H₂O₂ for 15 min. H₂O₂ was washed out and cells were collected after the indicated time. ROS levels were measured by flow cytometry. The ROS levels increase was calculated by subtracting ROS levels in untreated cells (measured in parallel). The mean and SD from 2 to 3 independent experiments are shown (after standardisation relative to 1 for untreated cells). (D,E) U2OS cells were transfected using the indicated siRNA and 48 hours following transfection, cells were subjected to comet assay. Representative cells are shown in (D). The mean and SD of the proportion of comet-positive cells (tail moment >5) from three independent experiments are shown in (E) (calculated relative to 100 for cells transfected with the p400-1 siRNA).