Fig. 6

Elevated levels of Crry expression and Akt activation in iNSC-derived neurons receiving CR2-Crry treatment. a, b Representative staining for the p-AKT⁺ (green, a), AKT⁺ (green, b), and Crry⁺ (red) depicted p-AKT, AKT, and Crry levels in neurons derived from iNSCs among the PBS (neurons receiving PBS treatment), CHI (neurons receiving CHI mouse serum treatment), and CHI+CR2-Crry (neurons receiving CHI mouse serum and CR2-Crry treatment) groups. Nuclei were counterstained with DAPI (blue). c, d Histograms showing the ratio of p-AKT⁺/Crry⁺ (c, the number of p-AKT and Crry double-positive cells/the total number of DAPI-positive cells) and AKT⁺/Crry⁺ (d, the number of AKT and Crry double-positive cells/the total number of DAPI-positive cells) cells among the three groups (n = 6/group; one-way ANOVA, *P < 0.05, **P < 0.01, ***P < 0.001). e, g, i Representative flow cytometric analysis of Crry (e), p-AKT (g), and AKT (i) expression in neurons derived from iNSCs among the PBS (blue line), CHI (green line), and CHI+CR2-Crry (red line) groups. Isotype antibodies were used as controls (black line). f, h, j Histograms indicating the median fluorescence intensity (MFI) values of Crry (f), p-AKT (h), and AKT (j) expression in iNSC-derived neurons among the three groups (n = 3/group; one-way ANOVA, *P < 0.05, **P < 0.01, ***P < 0.001). Scale bar = 100 μm (15 μm)