Figure 2

LPS induces PCT expression in HepG2 hepatocytes through NF-κB activation. (a) HepG2 cells were stimulated with LPS (5 μg/mL) for the indicated time. Cytoplasmic and nuclear extracts were assessed by Western blotting using specific antibodies against IκBα, p-IκBα and NF-κB p65, as well as β-actin and lamin A/C as loading controls for the cytoplasmic and nuclear fractions, respectively. Left, representative data of three independent experiments. Right, quantitative analysis of the band intensities of three independent experiments. Full-length blots are presented in Supplementary Fig. S1. (b,c) HepG2 cells were pretreated with PDTC (30 μg/mL) or Bay11-7082 (3 μg/mL) for 1 hr and stimulated with LPS (5 μg/mL) for 6 hr. The amount of intracellular PCT mRNA (b) and PCT protein in the supernatant (c) was detected, respectively. Data are represented as the mean ± SD of three experimental replicates. (d) HepG2 cells transiently transfected with NF-κB p65 siRNA were treated with LPS (5 μg/mL) for 6 hr. The level of PCT mRNA expression was detected by qPCR. Data are represented as the mean ± SD of three experimental replicates. ^*p < 0.05; ^**p < 0.01.