Figure 6

(A) Repin 1 mRNA expression of HepG2 cells 72 h after treatment with 0, 10, 20, 40, 80 and 160 nM siRNA specific for Luciferase (siLuci) and Repin1 (siRepin1) (n = 3–4 per group and concentration) and (B) proliferation of these cells using a BrdU proliferation assay (n = 9 per group and concentration). Values are given as means ± SEM (n = 4 per genotype and time point; *p < 0.0083 vs. LRep1+/+ of the individual time point, Mann-Whitney Rank Sum test followed by Bonferroni correction. (C) FACS analysis of Cd36 of primary isolated wildtype hepatocytes (Wt), which additionally were transfected with siLuci or siRepin1, and of hepatocytes of LRep1−/− mice under normal (left) and steatotic (right) culture conditions for a period of 72 h. Data are presented as box plots indicating the median, the interquartile range in form of a box and the minimum and maximum as whiskers. ^#p < 0.05 vs. Wt siLuci of the individual culture condition, Mann-Whitney Rank Sum test. (D) Repin1 mRNA expression of primary hepatocytes of the different genotypes and treatment groups as described above. *p < 0.01 vs. Wt; ^#p < 0.05 vs. Wt siLuci of the individual culture condition, Mann-Whitney Rank Sum test. (E) Oil Red O staining of primary isolated wildtype hepatocytes (Wt), which additionally were transfected with siLuci or siRepin1, and of hepatocytes of LRep1−/− mice under steatotic culture conditions for a period of 24 and 72 h. Scale bars represent 50 µm. (F) Scanning and electron microscopic images of primary isolated Wt hepatocytes (Wt), which additionally were transfected with a siRNA for Luciferase (siLuci) and Repin1 (siRepin1) as well as of LRep1−/− hepatocytes after 72 h of FA exposure. Compared to controls lipid droplets (white asterisks) appear smaller in Repin1-deficient hepatocytes. Scale bars represent 10 µm.