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. 2013 Jan 14;8(1):e53168. doi: 10.1371/journal.pone.0053168

Figure 3. DhL treatment causes the accumulation of DNA damage markers.

Figure 3

Unsynchronized HeLa cells were treated with 0, 20, or 30 µM DhL and lysed or fixed at the indicated times. (A) Immunoblot analysis of phospho-ATM (p-ATM) and phospho-Chk1 (p-Chk1). Representative assay of 3 independent experiments. β-actin was employed as a loading control. Cells were stained with DAPI to visualize the nuclei and treated with specific antibodies for γH2AX (B) and 53BP1 (C). Left: representative fields from 24 h treatment. Insets: magnification of the areas indicated by boxes in the top row. Representative fields for 48 h treatment are shown in Fig. S3. Right: quantification of the number of cells with more than 10 γH2AX foci and more than 5 53BP1 foci. At least 200 nuclei were scored for each sample. Bar: 10 µm. Data represent mean ± SEM of 3 independent experiments. * p≤0.05, ** p≤0.01 vs. control group (0 µM DhL).