Fig. 1

PNS promoted primary mouse hepatocyte proliferation via the PI3K/AKT/mTOR signaling pathway. The primary mouse hepatocytes were treated with a range of concentrations of PNS for 24 h. Cells were preincubated with or without 10 μM LY294002 for 1 h and were then treated with 0.12 mg/ml PNS for 24 h. (a) The cell viability was determined by CCK-8 Kits. (b) Primary mouse hepatocytes were pretreated with 0.00, 0.06, 0.12 and 0.25 mg/ml PNS for 24 h and EdU (pink) staining was measured (magnification: × 400, Scale bars represent 50 μm). (c) Quantification of EdU-positive cells. (d) Representative immunofluorescent images of primary mouse hepatocyte proliferation measured by EdU staining (magnification: × 400, Scale bars represent 50 μm). (e) Quantification of EdU-positive cells. (f) Total and phosphorylated protein levels of PI3K, AKT and mTOR in primary mouse hepatocytes. Values represent the mean ± SEM of 3 independent experiments. (g)(h)(i) Bar graph shows the quantification of total and phosphorylated protein levels of PI3K, AKT and mTOR. Values represent the mean ± SD of at least three independent experiments. **P < 0.01, *** P < 0.001