Figure 6

17α-E2 suppresses SCD1 expression in hepatocytes and HSCs in vitro. (A) HepG2 cell viability [n = 3/treatment/timepoint] and (B) HepG2 Bax mRNA [n = 3/treatment/timepoint] following 6, 12, and 24 h of treatment with VEH, VEH + PA (0.5 mM), or VEH + PA + 17α-E2 (100 nM, 10 nM, 1 nM). (C) LX-2 cell viability [n = 3/treatment/timepoint] and (D) LX-2 Col1a1 mRNA [n = 3/treatment/timepoint] following 6, 12, and 24 h of treatment with VEH, VEH + TGF-ꞵ1 (5 ng/ml), or VEH + TGF-ꞵ1 + 17α-E2. (E) HepG2 Scd1 mRNA [n = 3/treatment/timepoint] following 6 and 12 h of treatment with VEH, VEH + PA, or VEH + PA + 17α-E2. (F) HepG2 SCD1 protein [n = 4/treatment] following 12 h of treatment with VEH, VEH + PA, or VEH + PA + 17α-E2. (G) Representative immunoblots of SCD1 and GAPDH in HepG2 cells following 12 h of treatment with VEH, VEH + PA, or VEH + PA + 17α-E2. (H) LX-2 Scd1 mRNA [n = 3/treatment/timepoint] following 12 and 24 h of treatment with VEH, VEH + TGF-ꞵ1, or VEH + TGF-ꞵ1 + 17α-E2. (I) LX-2 SCD1 protein [n = 4/treatment] following 12 h of treatment with VEH, VEH + TGF-ꞵ1, or VEH + TGF-ꞵ1 + 17α-E2. (J) Representative immunoblots of SCD1 and GAPDH in LX2 cells following 12 h of treatment with VEH, VEH + TGF-ꞵ1, or VEH + TGF-ꞵ1 + 17α-E2. Green dotted lines in panels (A) and (C) represent 100% viability, whereas the red dash lines represent a 40% loss of viability in panel (A) and a 40% gain in viability in panel (C). All data are presented as mean ± SEM and were analyzed within timepoint by one-way ANOVA (C–F,H,I) with Tukey post-hoc comparisons. Statistics were not performed on data shown in panels (A) and (C). *p < 0.05. We did not indicate statistical differences between VEH and 17α-E2 treatment groups for purposes of visual clarity.