Fig. 3
Effects of DNLS inhibitors on induction of APOA1 RNA expression and on cellular lipid concentration in cells treated with ethanol. Relative RNA expression levels were determined by RT-qPCR as described in Methods. a and b Expression of APOA1, LDLR, HMGCR and SREBF2 in HepG2 cells cultured in LPDS medium and treated at the indicated final concentration of ethanol (100 mM) and/or atorvastatin (20uM). c HepG2 cells were cultured in standard medium, and exposed to combinations of 100 mM ethanaol, 20 uM atorvastatin, and/or 20 uM TOFA as per Materials and Methods. The error bars represent the SD from the mean of five assays of an individual experiment. *Significant difference, ethanol treatment versus untreated control (1 way ANOVA with multiple comparisons, n = 5; *P < 0.05, **P < 0.01, ***P < 0.001 vs. control). For the effect of DNLS Inhibitors on cellular lipid concentration. HepG2 cells were cultured 21 days in DMEM+ 10% FBS for differentiation, after O/N starvation, cells were incubated with [14C] oleic acid and treated with atorvastatin (20 uM) or ETOH (100 m M) + Atorvastatin (20 mM) for 24 h (d and e). For the f cells were treated with TOFA (20 uM) or ETOH (100 m M) + TOFA (20 mM) for 24 h with the concentration indicated of ETOH. After the incubation, lipids were extracted, separated by thin-layer chromatography and quantified as described in Materials and Methods. Data were analyzed as dpm/mg of total protein and represent means±SD for n = 3 independent experiments. *P < 0.05, **P < 0.01, ***P < 0.001 vs. control