Figure 2.

Effects of Supernatant from HCV-Replicating Cells on miR-192 Expression in HSCs

(A) Experimental scheme (left). LX-2 cells were treated with supernatant from naive Huh-7 or JFH-1 stable cells for 3 days. The RNA levels of COL1A1, α-SMA, and TGF-β1 and miR-192 levels in LX-2 cells treated with JFH-1 supernatant were quantified relative to those in cells treated with naive Huh-7 supernatant (right). (B) Effects of TGF-β1 on miR-192 expression in HSCs are shown. LX-2 cells were cultured with serum-free medium for 16 h and then treated with different concentrations of recombinant TGF-β1 protein for 72 h. The mRNA levels of COL1A1, α-SMA, and TGF-β1 and miR-192 levels in LX-2 cells were assessed relative to those in control PBS-treated cells. (C) Effects of miR-192 introduction or depletion on Huh-7 cells are shown. Experimental scheme is shown (left). Huh-7 cells were transfected with miR-192 mimic RNA or anti-miR-192 for 48 h and then the supernatant was collected and used to treat LX-2 cells for 72 h. The miR-192 copy number in each supernatant was quantified (middle). miR-192 levels in LX-2 cells treated with each type of supernatant were assessed relative to those in cells treated with supernatant from negative-control miRNA (siNTC)- or scramble RNA (scr)-treated Huh-7 cells (right). All data are presented as the means of at least three independent experiments, each performed in triplicate. Error bars represent the SEM. p values were determined via a one-tailed unpaired Student’s t test. *p < 0.05; **p < 0.01; ***p < 0.001; n.s., non-significant.