Fig. 7: RPV’s effect on cell viability is partially rescued by silencing proteins involved in autophagy.
From: Implication of autophagy in the antifibrogenic effect of Rilpivirine: when more is less
Specific autophagy genes were silenced by siRNA in LX-2 cells (24 h) which were then treated with TGF-β or TGF-β plus different concentrations of RPV for 48 h. Representative WB images and quantification of the protein levels of autophagy related genes—(A) SQSTM1, (B) ATG5 and (C) BECN1 following transfection with siRNA (D) Representative WB images of ATG5 and collagen 1A1 in control or ATG5-silenced cells. E Representative WB images of p62 and collagen 1A1 in control or SQSTM1-silenced cells. GAPDH was employed as a loading control. Viability of cells exposed to control siRNA vs (F) ATG5, (G) BECN1 and (H) SQSTM1 silenced cells treated either with TGF-β or different concentrations of RPV plus TGF-β for 48 h. Cell viability was assessed using acid phosphatase assay. All data represent mean ± SEM (n = 3) and are expressed as in relation to those of untreated cells in each experiment, which was considered 100% (both in control siRNA and in gene-specific siRNA). Statistical analysis was by two-way ANOVA with Bonferroni post-test of control siRNA vs gene-specific siRNA (*, ** and **** represent a p value < 0.05, < 0.01 and < 0.0001, respectively).
