Fig. 1

ACLY and ME1 are activated by TNFα in primary human hepatocytes. Primary human hepatocytes (HH) were treated with 5 ng/mL TNFα alone (TNFα) or combined with 500 μM HCA (TNFα + HCA). Unstimulated cells (C) were used as a negative control. Real time PCR experiments were performed to evaluate ACLY (A) and ME1 (D) mRNA levels. Data are representative of 3 independent experiments and are presented as mean ± SD (error bars). Western blot analyses revealed ACLY (B) and ME1 (E) protein content. Immunoblot data are representative of at least 3 independent experiments. Values obtained after normalization to β-actin are reported under western blot images. Protein expression levels in untreated HH (C) were taken as 1. C ACLY activity in human hepatocytes. F Representative photomicrographs of intracellular lipids staining. Photographs typical of those taken three separate experiments are shown. G Quantitative assessment of Oil Red O staining. Data are shown as mean ± SD (error bars) and derived from 3 experiments. In A, C, D differences were significant according to Student’s t-test (*p < 0.05 and ***p < 0.001). Statistical significance of difference was evaluated in G by the mean of one-way ANOVA followed by Tukey’s multiple comparison test and different letters indicate significant differences between treatments at p < 0.05