Fig. 3

Plin5 stimulated lipid accumulation and reduced the lipolysis induced by NS5A. a Oil red O (ORO) staining of liver tissues from wild-type (Plin5^+/+) and Plin5-null (Plin5^−/−) mice. Scale bar = 50 μm. b & c Triglyceride (TG) and non-esterified fatty acid (NEFA) content was quantified by commercial kits after extraction with hexane/isopropanol. d Primary hepatocytes were isolated from wild-type and Plin5-null mice and then infected with control (Ad-Null) and NS5A-expressing (Ad-NS5A) adenoviruses. The lipid droplets (LDs) in primary hepatocytes were stained with BODIPY 493/503 and observed under a fluorescence microscope. Scale bar = 20 μm. e After extraction with hexane/isopropanol, the TG content was quantified in wild-type (Plin5^+/+) and Plin5-null (Plin5^−/−) hepatocytes. f Primary hepatocytes with or without NS5A expression were treated with triacsin C (5 μM), an inhibitor of acyl-CoA synthase, and the NEFA content was determined over 8 h. g Wild-type (Plin5^+/+) and Plin5-null (Plin5^−/−) primary hepatocytes with or without NS5A expression were incubated with 200 μM OA for 24 h. The cells were then treated with 5 μM triacsin C to inhibit further triacylglycerol synthesis. Cellular lipids were extracted with hexane/isopropanol, and then the TG content was quantified. *P < 0.05, **P <0.01, ***P < 0.001. Data are means ± SEs for triplicate samples for each representative experiment