Fig. 5

FSZ NPs promote neural differentiation through the WNT4 signaling pathway. MSCs were treated with FSZ (0 and 40 μg/ml) for 6 days in neural inducing medium, total mRNA was extracted and subjected to RNA-sequence assay for gene expression identifying. (A) Volcano maps and (B) heat maps are displayed. (C) GO and (D) KEGG analyzed the up-regulated genes associated with FSZ. (E) MSCs were treated with FSZ (0, 5, 10, 20, 40 and 80 μg/ml) for 6 days in neural inducing medium, Western blot assay was used to detect the expression of WNT4, Axin2, GSK3β, p-GSK3β, β-catenin, active-β-catenin, JNK1/2, p-JNK1/2, P38, p-P38 and GAPDH. (F) MSCs were treated with FSZ (0 and 40 μg/ml) combination with transfection of WNT4-siRNA1 or WNT4-siRNA2 for 6 days in neural inducing medium, Western blot assay was used to detect the expression of WNT4, GSK3β, p-GSK3β, β-catenin, active-β-catenin, Tuj1, MAP2 and GAPDH. Protein levels were quantified using ImageJ (1.53a for Mac). Data are presented as means ± SD (n = 3). Statistical analysis was performed using one-way ANOVA. # P < 0.05, compared with FSZ group; * P < 0.05 compared with Con group