Skip to main content
. 2019 Mar 8;11(3):334. doi: 10.3390/cancers11030334

Figure 6.

Figure 6

Effects of suppressing hnRNP A1 and/or A2 on DDR in normal fibroblasts and hTERT-immortalized normal fibroblasts. (A) Normal fibroblasts (HFF3, MRC5, and HFB) and hTERT-immortalized normal fibroblasts (HFF3-hTERT, MRC5-hTERT, and HFB-hTERT) were transfected with siRNA targeting hnRNPA1 (siA1), hnRNPA2 (siA2), both hnRNP A1 and A2 (siA1/A2), or with a non-targeting sequence (siNT). After 72 h, cell lysates were analyzed for expression levels of hnRNP A1, hnRNP A2, and γH2AX by Western blotting. β-Actin served as a loading control, and cells treated with 50 μM etoposide (Etop) for 12 h served as a positive control; (B) Transfected HFF3-hTERT cells were fixed and immunostained for γH2AX (red) and TRF2 (green), and nuclei were counterstained with DAPI (blue). HFF3-hTERT cells treated with 50 μM etoposide (Etop) for 12 h were included as a control; the scale bar equals 5 μm. (C) The percentage of nuclei positive for γH2AX staining was determined from analysis of ~100 nuclei from each experiment; the data shown in the left panel are from 2 independent experiments. The percentage of γH2AX co-localized with TRF2 was determined from analysis of 50 γH2AX -positive nuclei from each experiment; the results shown in the right panel are from 2 independent experiments. * p < 0.05 and *** p < 0.001 versus siNT control.