Figure 6

Cholesterol provoked a proinflammatory response in Kupffer cells and increased apoptosis in hepatocytes. (A,B) Primary mouse Kupffer cells were stimulated with 1 mg/mL cholesterol crystals (Cho-C) for 8 h. Relative mRNA expression levels of (A) the immune cell-recruiting chemokines Ccl2 and Cxcl2 and (B) the proinflammatory cytokines tumor necrosis factor α (Tnf) and oncostatin M (Osm) were quantified by RT-qPCR with β-actin (Actb) as reference gene. (C) Primary mouse hepatocytes were stimulated with 25 µg/mL cholesterol crystals (Cho-C), 1 ng/mL TNFa and 300 nM actinomycin D for 8 h. Apoptosis was determined by caspase 3 activity measured with a fluorescence-based assay and normalized to protein content. Values represent means ± standard error of the mean of a minimum of three independent experiments. Statistics: (A,B) Two-sided Student t test for unpaired samples, *p <0.05. (C) Two-sided Student t test for unpaired samples or, where appropriate, two-way analysis of variance with Tukey’s post hoc test for multiple comparisons, *p <0.05.