Figure 4

MIR181A1HGkd cells show increased DNA damage as assessed by 53BP1 and γH2AX staining. (A) Immunofluorescence microscopy of hMSC-hTERT20 cells, CRISPRi control and MIR181A1HGkd for 53BP1 (red) and γH2AX (green). Nuclei were counterstained with DAPI (blue). 53BP1 and γH2AX are diffuse in the nucleoplasm in undamaged cells and upon damage/dsDNA breaks, shows punctate focal staining patterns. Cells with high γH2AX foci are highlighted by *. Scale bars represent 20 µm. Quantification of the number of (B) γH2AX and (C) 53BP1 foci per nuclei was made using Volocity. For each cell line, > 700 cells were analyzed from 2 independent experiments. *p < 0.05; ***p < 0.005. Data are presented as mean ± SD.