Fig. 4.

Enhanced levels of Crry expression and Akt activation in iNSC-derived neurons receiving astrocyte culture supernatants. a, b RT-QPCR was utilized to determine the expression of the Crry (a) and Akt (b) genes in neurons derived from iNSCs among the four sub-groups, which were separately treated as follows: (i) CHI mouse serum diluted (20%) in DMEM/F12 (1:1), (ii) CHI mouse serum diluted (20%) in the astrocyte culture supernatants, (iii) CHI mouse serum diluted (20%) in DMEM/F12 (1:1) containing purified rat anti-mouse Crry antibody at 5 μg ml⁻¹, and (iv) CHI mouse serum diluted (20%) in the astrocyte culture supernatants containing purified rat anti-mouse Crry antibody at 5 μg ml⁻¹ for 45 min at 37 °C (n = 3/group; one-way ANOVA, *P < 0.05, **P < 0.01, ***P < 0.001 versus i, iii, and iv sub-groups, respectively). c Representative immunoblots depicting the levels of Crry, p-AKT, and AKT in neurons derived from iNSCs among the four sub-groups. d–g Histograms showing the relative levels of Crry (d), p-AKT (e), AKT (f), and p-AKT/AKT (g) in neurons derived from iNSCs among the four sub-groups (n = 6/group; one-way ANOVA, **P < 0.01, ***P < 0.001 versus i, iii, and iv sub-groups, respectively)