Figure 4.

Primary HSCs Activation by Exosome Isolated from HCV-Replicating Hepatocytes

(A–C) Quantification of fibrotic marker RNAs and proteins in primary HSCs treated with each type of exosomes. (A) Relative intracellular mRNA levels of fibrosis markers and (B) copy numbers of miR-192, miR-122, and miR-19-3p in primary HSCs treated with each type of exosome are shown. (C) Immunoblot analysis of the fibrosis markers COL1A1 and α-SMA, as well as TGF-β1, in primary HSCs treated with each type of exosome (left) and quantification of their levels relative to those of tubulin (right) are shown. (D) Effects of exosomes on primary HSCs activation are shown. Primary HSCs were treated with each exosome type for 72 h, fixed, stained, and visualized using fluorescence microscopy. Representative images (left) of α-SMA and DAPI staining in primary HSCs are shown. Scale bars represent 50 μm. α-SMA protein levels (right) in primary HSCs treated with each type of exosomes were quantified from more than five fields and shown relative to the levels in cells treated with Huh-7 exo. Data are also shown in Figure S5. 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 using a one-tailed unpaired Student’s t test. *p < 0.05; **p < 0.01; ***p < 0.001; n.s., non-significant.