Elevated spontaneous DNA damage in PDAC cells.

<p>(A) Western blot analysis of γH2AX in HPDE and 18 PDAC cell lines. Samples from 8988T and Panc1 irradiated at 4Gy were used as positive controls and actin served as the loading control. Experiments were done at least three times. (B) Quantitation of γH2AX expression by densitometry was normalized to actin expression and presented as relative to HPDE. Data is shown from three independent experiments with error bars representing standard deviations. (C) Immunofluoresence for basal γH2AX foci in three representative cell lines (HPDE, 8988T and HPAC). Green: γH2AX; Blue: DAPI (nucleus). Scale bar equals 10 µm. Quantitation was performed in (D) and shown as percentage of cells with more than 5 foci. Experiments were done three times and average values were calculated. Error bars represent standard deviation from three individual experiments. (E) Neutral comet assay was performed to directly assess DNA double strand breaks and the data expressed as tail moment (Tail moment = tail length ×% of DNA in the tail). For all panels, asterisks show a statistically significant increase as compared to HPDE by t-test (p≤0.05).