Fig. 4

Effect of ASD/XLMR associated NL4X variants on the synaptogenic activity and the protein metabolism. a Quantitative result of synaptogenic activity of NL4X variants. Intensity of VGAT puncta was counted and standardized by the area of HEK293 cells (n = 12–17, ***p < 0.001 vs WT by Tukey-Kramer multiple comparisons test). b Representative immunoblot of cell lysates of COS-1 cells transfected with missense variants of NL4X. c Quantitative analysis of the ratio of mature NL4X (n = 3, ***p < 0.001 vs WT by one-way ANOVA followed by Dunnett test; n.s., not significant). d Representative immunoblot of biotinylated cell surface NL4X variants. e Quantitative analysis of the western blot for biotinylated WT and variants (n = 3, *p < 0.05, **p < 0.01, ***p < 0.001 vs WT by one-way ANOVA followed by Dunnett test; n.s. not significant). f Representative immunoblot of conditioned medium from COS-1 cells transfected with missense variants of NL4X. g Quantitative analysis of the western blot for sNL4X (n = 3, *p < 0.05, **p < 0.01, ***p < 0.001 vs WT, by one-way ANOVA followed by Dunnett test; n.s., not significant)