FIG 2.

Effects of ATM, ATR, and CHK1 inhibitors on CCC DNA formation during HBV infection in HepG2-NTCP cells. The HepG2-NTCP cells were infected with the HBV inoculum harvested from HepAD38 cells. The following inhibitors were added at the indicated concentrations at the same time as the inoculum and maintained until the cells were harvested: the ATM inhibitors (ATM-i) KU-55933 and KU-60019; the ATM/ATR dual inhibitors (ATM/ATR-i) CGK733 and Torin2; the ATR (ATR-i) inhibitors AZD6738 and VE-821; and the CHK1 inhibitor (CHK1-i) CHIR-124. HBV PF DNA was extracted from the infected cells at 3 days postinfection and measured by Southern blotting using a ³²P-labeled HBV DNA probe. (A and B) Representative Southern blot autoradiograms. The brackets indicate the putative RC DNA processing products accumulating under conditions of ATR-CHK1 inhibition (see Fig. 8, below). (C). Quantitative analysis of Southern blot results from multiple independent experiments. The data are expressed as CCC DNA levels normalized to those of PF-RC DNA, with the normalized CCC DNA level from the mock-treated cells set to 1.0. *, P ≤ 0.05; **, P ≤ 0.01; ***, P ≤ 0.001. The vertical thin lines in the images denote where the different parts of the same gel, with the same exposure, were spliced together in order to remove other parts of the gel that are not presented here.