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. 2016 Feb 8;7(11):12682–12692. doi: 10.18632/oncotarget.7259

Figure 3. As2O3-induced DNA damage and chromosome instability is associated with degradation of telomeric G-overhang.

Figure 3

A. Representative immunofluorescence images of ATR, 53BP1, γ-H2AX and Mer11 foci in U87 cells treated with 4 μM As2O3 for 48 h. Scale bar = 15 μm. B. Immunoblots showing up-regulation of p-ATM, ATR, 53BP1, γ-H2AX, p21 and Mer11 proteins in U87 cells treated for 48 h with 4μM As2O3. Immunoblotting β-actin confirmed equivalent protein loading. Each experiment was repeated three times. C. Telomere fusion induced by treatment with 4 μM As2O3 for 48 h. Chromosomes were stained with Giemsa. Scale bar = 5 μm. D. Hybridization protection assays (HPAs) were performed on genomic DNA isolated from glioma cells treated with As2O3 to assess G-overhang length and total telomere length. Exo 1 nuclease digestion was used to assess integrity of the 3′-overhang. Luminescence intensity in arbitrary units (AU) was normalized against Alu probe. The mean of three independent experiments with comparable results is shown. Error bars indicate ± s.d., **P < 0.01, two-tailed Student's t-test.