Figure 5.

Nestin depletion suppresses DNA damage repair and increases IR sensitivity. (a) Nestin-depleted (NESi) or control (NC) 5-8F cells were treated with the indicated concentrations of doxorubicin (Dox). After 24 h, Dox was removed and cells were cultured in fresh medium for the indicated time periods. The protein levels of γ-H2AX were examined by western blotting. β-Actin was tested as a loading control. (b) Quantification of the levels ofγ-H2AX protein from panel a. Values were normalized to β-actin. (c) 5-8F/NC and 5-8F/NESi cells were exposed to 4 Gy of IR, and subjected to alkaline comet analysis at the indicated time points. Magnification: × 100. (d) Quantification of tail moments from panel c. Tail moments for each condition was calculated on a minimum of 100 cells for each data point