FIG 2.

Transfection of pIHBoV1 induces a DDR in HEK293 cells. HEK293 cells were transfected with the HBoV1 infectious clone pIHBoV1 or its backbone plasmid, pBB, and subjected to IF and Western blot analyses at the indicated times posttransfection. HU-treated cells were used as a DDR-positive control. (A and B) IF analysis. Cells were stained with anti-NS1C and anti-γH2AX antibodies (A) and with anti-NS1C and anti-p-RPA32 antibodies (B). Nuclei were stained with DAPI. Confocal images were taken at a ×100 magnification. (C and D) Western blot analysis. (C) The cells were analyzed by Western blotting using the antibodies anti-γH2AX and anti-p-RPA32. β-Actin served as a loading control. (D) The levels of γH2AX and p-RPA32 expression in each group were quantified, and the levels relative to those in HU-treated cells were calculated and plotted. Averages and standard deviations obtained from three independent experiments are shown.