Figure 2
R-III treatment and albumin expression promote the nuclear translocation of NF-κB. (A, B) Hepatic stellate cells after passage 1 (HSCs-P1) were treated with HPLC-purified, His-tagged R-III (0.25 μM) or transfected with plasmids encoding His-tagged, wild-type or mutant albumin (R410A, Y411A, K525A) and then analyzed by immunofluorescence using the anti-p65 antibody (A). The cell nuclei were stained with DAPI (blue). (B) Nuclear and cytoplasmic fractions were prepared from the treated cells and subjected to western blotting using anti-NF-κB antibody. Lamin A/C and α-tubulin were used as internal controls. Bar graph represents the average of two independent experiments. Full-length blots are presented in Supplementary Fig. S4. (C) Total RNA was isolated from the HSCs on days 2, 3, and 4 after seeding (different stages of activation), and HSCs-P1 with or without R-III treatment, and the expression levels of IL-1α, IL-1β, IL-6, and TNF-α were then analyzed by real-time PCR. *P < 0.05, paired t-test (n = 3) (compared with untreated HSCs-P1).
