Figure 2
IR of p27KO cells is associated to high residual DNA damage and micronuclei frequency. (a) Western Blot analysis of phospho-Ser139-Histone 2AX (γH2AX) in lysates from indicated cell clones, non-irradiated (NIR) or irradiated with 1, 2 and 10 Gy and harvested after 30 minutes or 24 hours. Tubulin was used as loading control. The bottom panel shows p27 expression levels in the different cell clones. On the right, quantification of normalized levels of γH2AX is reported. (b,c) Graphs report the quantification of the γH2AX positive foci/cell (b) and the γH2AX positive cells (c), at 24 hours after 2 Gy, normalized for the number of the corresponding NIR and expressed as mean of three independent experiments performed in duplicates, with SD. (d) Graph reports the frequency of micronuclei positivity found at 24 hours in NIR cells and 2 Gy irradiated cells expressed as percentage of positive cells over the total cell number. (e) Representative pictures of NIR cells (left panels) or 2 Gy irradiated cells after 24 hours (right panels), showing the presence of micronuclei (yellow arrows). Cells were stained with DAPI. *Indicates a p ≤ 0.05; **p ≤ 0.01.
