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. Author manuscript; available in PMC: 2018 Nov 1.
Published in final edited form as: Alcohol Clin Exp Res. 2017 Oct 11;41(11):1875–1885. doi: 10.1111/acer.13496

Figure 4. In vitro inhibition of miR-21 restored Smad7 and partially attenuated TGFβ1 protein expression in alcohol-treated NIH 3T3 fibroblasts.

Figure 4

NIH 3T3 fibroblasts were transfected with miR-21 inhibitor (miR-21 inh; 10μM) or anti-miR negative control (scramble) and treated ± alcohol (Alc; 60 mM) starting at 24 hours after transfection. At 72 hours, cells were collected for protein expression analyses by Western immunoblot. (A) Consistent with the findings in PLFs, alcohol-treated NIH 3T3 cells transfected with negative control showed a decrease in Smad7 protein expression. Inhibition of miR-21 attenuated alcohol-mediated Smad7 suppression (normalized to GAPDH; representative gels shown above summary data). (B) Similarly, alcohol-treated NIH 3T3 cells transfected with negative control showed a significant increase in both latent and active TGFβ1 protein expression. Inhibition of miR-21 partially attenuated alcohol-induced latent and active TGFβ1 protein expression (normalized to GAPDH; representative gels shown above summary data). *P<0.05 compared to negative control (scramble RNA) transfected group. **P<0.05 compared to negative control (scramble RNA) + alcohol treatment group.