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. 2016 Mar 22;6:23506. doi: 10.1038/srep23506

Figure 4. BRE is responsive to DNA damage.

Figure 4

(a) Viability of early passage WT and BRE−/− fibroblasts in response to a dose range of gamma irradiation. Percentage cell viability was determined using CCK-8 assay on day 7 after each indicated irradiation dose. Shown here is the representative result of fibroblasts at passage number 4. Data represent the mean ± SD of thee independent experiments. (b) BRE mRNA levels at indicated time points after 10 Gy of gamma irradiation as determined by RT-qPCR. Data represent the mean ± SD of triplicate wells. **denotes P-value < 0.01, and ***denotes P-value < 0.001 for the difference compared with 0 h. (c) Immunofluorescence staining with anti-BRE antibody (green) and anti-γ-H2AX antibody (red), together with DAPI nuclear staining (blue), of WT fibroblasts at 24 h after 10 Gy of gamma irradiation compared with the control without irradiation. Each panel comprises 4 images of the same field of cells showing the staining by DAPI (top left), γ-H2AX (bottom left), BRE (top right) and the composite of γ-H2AX and BRE (bottom right). Arrowheads indicate examples of increased nuclear BRE expression in association with DNA damage. (d) Western blot analysis for BRE expression of WT fibroblast whole cell lysates at indicated time points after 10 Gy of gamma irradiation. Relative expression levels of BRE versus α-tubulin are shown as percentage values.