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. 2016 Jun 21;7(32):51965–51980. doi: 10.18632/oncotarget.10207

Figure 3. Suppression of miR-99a and 100 promotes DNA repair enhance recruitment of DNA repair proteins.

Figure 3

A. Quantification of positive nuclear phospho-ATM/ATR substrate and phospho-P53 (s-20) stained CB cells transfected with miR-99a and 100 inhibitor. Immunofluorescence staining was performed was performed 30 minutes after exposure to 5-Gy (n=3 PCa, each sample in triplicate), >250 cells/sample were counted. B. Quantification of γH2AX immunofluorescence foci/nucleus at multiple time points after transfection of miR-99a/100 inhibitor in CB cells following 5-Gy radiation exposure (n=3 PCa, each sample in triplicate), >250 cells/sample were counted. Line represents 50% of total γH2AX foci/cell. C. Quantification of nuclear pan-histone 3-acetylation immunofluorescence staining intensity by Velocity Quantitation software in miR-99a and 100-inhibitor transfected CB cells 30 minutes after exposure to 5-Gy radiation (n=3 PCa). n indicates total number of cells included in the analysis. D. Quantification of miR-inhibitor transfected CB cells exhibiting nuclear BRCA1 and RAD51 immunofluorescence foci, 120 minutes after exposure to 5-Gy radiation (n = 3 PCa, each sample in triplicate), >250 cells/sample were counted. E. Representative pictures of immunofluorescence staining for phosphop53 (s-20), cleaved caspase 3, and cleaved PARP expression in miR-99a/100 inhibitor transfected CB cells, 24 h after exposure to 5-Gy radiation (n=3 PCa, each sample in triplicate). The right panel shows quantitation of staining using Velocity Quantitation software. n indicates total number of cells quantified. Scale bar: 120 μm. Data are expressed as mean ± s.d. *P < 0.05, **P < 0.01, ***P < 0.001 (Student's ttest).