Fig. 4

Apelin and ML221 promoted differentiation of NSCs. A Expression of Apelin (green) in NSCs (scale bar = 120 µm). B Differentiated cell types were identified by expression of specific markers via immunofluorescence of GFAP (astrocytes, red) and NF200 (neurons, green) under different dose of Apelin and ML221 (scale bar = 120 µm). C A statistic summary of NeuN+ and GFAP+ cells proportions (***p < 0.001 NF200 vs. control group, ^###p < 0.001 GFAP vs. control group, mean ± SD). D, E RT-PCR results indicating different types of cells differentiated from NSCs following induction by Apelin and ML221. Primers for two targets were used: GFAP (astrocytes) and β-III-tubulin (neurons); a specific primer for GAPDH was also included in reactions (**p < 0.01, ***p < 0.001 vs. control group, mean ± SD)