Fig. 2

Metformin enhances NSCs proliferation and neural differentiation via AMPK-dependent regulation. (A, B) Representative western blot images of p-AMPK and AMPK and quantitative analysis of p-AMPK / AMPK expression level of NSCs with different treatments (N = 3). The relative expression level of target proteins was normalized by GAPDH and then calculated as fold changes of the control group. (C) Cell viability assay of NSCs with different treatments (N = 3). Cells were cultured in NSCs-specific growth medium for 24 h before assessment. The cell viability was calculated as fold changes of the control group. (D, E) Representative immunofluorescence images of EdU (red) positive cells and quantitative analysis of the proportions of EdU positive cells in NSCs (N = 15). Cells were cultured in NSCs-specific growth medium for 24 h before assessment. Cell nuclei were stained with Hoechst (blue). Scale bar = 50 μm. (F-H) Representative immunofluorescence images of NSCs double-labeled with Tuj-1 (red) and GFAP (green), and quantitative analysis of the respective proportions of Tuj-1 positive cells and GFAP positive cells in NSCs (N = 14). Cells were cultured in NSCs differentiation medium for 7 d before assessment. Cell nuclei were stained with DAPI (blue). Scale bar = 50 μm. All analyses were performed using one-way ANOVA and Dunnett’s post-hoc test. Data are shown as mean ± SEM. (* p < 0.05, ** p < 0.01, *** p < 0.001)