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. 2015 Jun 12;5:10867. doi: 10.1038/srep10867

Figure 5. Cav-1 knockdown sensitizes tumor cells to radiation by increasing mitotic catastrophe and attenuating DNA damage response signaling.

Figure 5

A. Quantification of the percentage of cells with mitotic catastrophe after 3 Gy radiation (left panels, *p < 0.05). Representative immunofluorescence images of presence or absence of mitotic catastrophe at 72 hours in scrambled control and siCav-1 cells (right panels). B. Quantification of average number of nuclear pH2A.X foci per cell at multiple time points (*p < 0.05). Representative immunofluorescence staining for pH2A.X foci after 3 Gy radiation at 24 hours in scrambled control siRNA and siCav-1 cells (right panels). C. Top - Quantification of average number of nuclear BRCA1 foci per cell at multiple time points (left; *p < 0.05). Representative immunofluorescence staining of BRCA1 foci after radiation at 24 hrs in scrambled control siRNA vs siCav-1 (right panels). Bottom - Quantification of average number of nuclear pDNA-PK foci per cell (left; *p < 0.05). Representative immunofluorescence staining of pDNA-PK foci after radiation at 24 hrs in scrambled control siRNA-and siCav-1 (right panels). D. Immunoblotting of scrambled control and siCav-1 treated cells with or without radiation at different time points for expression of phosphorylated BRCA1, total BRCA1, phosphorylated DNAPK, total DNAPK, and beta actin as control (top panel). Note the reductions in phospho-BRCA1 and phospho-DNA-PK at 24 hrs after radiation with Cav-1 depletion. Loss of Cav-1 also increases the degree of radiation-induced activation of apoptotic signaling at 24 hours after 3Gy (bottom panel). For nuclear foci and mitotic catastrophe experiments, at least 50 cells were counted per datapoint.