Figure 3.

TGF-β1 Elevates miR-96 Expression through Post-transcriptional Regulation

(A) qRT-PCR analysis of miR-96 expression in isolated hepatocytes, HSCs, and Kupffer cells from mouse livers at 0 and 50 dpi, and the miR-96 expression in HSCs of the wild-type mice at 0 and 50 dpi. (B) qRT-PCR analysis of tgf-β1 and il13 expression in the HSCs of infected mice. (C) qRT-PCR analysis of miR-96 and miR-214 expression in primary HSCs treated in vitro for 24 hr with IL10 (25 ng/mL), interferon (IFN)-γ (25 ng/mL), TGF-β1 (100 ng/mL), and IL13 (200 ng/mL). (D) qRT-PCR analysis of miR-96 expression in primary HSCs treated with different concentrations of TGF-β1 as indicated. (E) qRT-PCR analysis of miR-96 expression in primary HSCs treated with TGF-β1 (8 ng/mL) for various times. (F) qRT-PCR analysis of pri-miR-96 and pre-miR-96 expression levels in isolated HSCs from mice at 0 and 50 dpi. (G) qRT-PCR analysis of pri-miR-96 and pre-miR-96 expression in primary HSCs treated with TGF-β1 (10 ng/mL) for various times. (H) RNA-binding protein immunoprecipitation (RIP) analysis of post-transcriptional processing of miR-96. The cell lysates of HSCs isolated from infected (50 dpi) and uninfected (0 dpi) mice were immunoprecipitated with anti-SMAD2+SMAD3, anti-DROSHA, or normal rabbit IgG and subjected to qRT-PCR analysis. (I) In vitro RIP analysis of post-transcriptional processing of miR-96 and miR-21 maturation. qRT-PCR analysis of miR-96 and miR-21 expression in primary HSCs treated with TGF-β1 (100 ng/mL) or IL13 (200 ng/mL) for 24 hr. HSC lysates were immunoprecipitated and subjected to qRT-PCR analysis as described above. Data are represented as mean ± SD. *p < 0.05, **p < 0.01, ***p < 0.001 compared between indicated groups.