Fig. 4

Silencing USP18 in HepAD38 suppressed HBV production. HepAD38 cells were seeded at 3 × 10⁵/ml, 2 ml per well in 6-well plates. 24 h later, the cells were left untreated or transfected with 20 nM siUSP18 or 20 nM NC. 48 h post transfection, knockdown efficiency was evaluated by detecting USP18 mRNA expression (a, left), which was further confirmed by western blot (a, right). Intracellular total RNA and DNA, as well as supernatant DNA, were collected respectively. Real-time PCR was performed to detect supernatant HBV total DNA (b), intracellular HBV total DNA (c), HBV cccDNA (d) and pgRNA (e). siUSP18, USP18 small inhibitory RNA; NC, the negative control siRNA. Results are presented as means ± SD (n ≥ 3). *p ≤ 0.05; **p ≤ 0.01; ***p ≤ 0.001