1. Fig. 5. Nutlin-3 treatment rescues the proliferation and differentiation of NPCs in vitro.
Nutlin-3 treatment rescues the proliferation and differentiation of NPCs in vitro. (A) Schematic showing that Nutlin-3 may inhibit the interaction between p-MDM2(Ser166/Ser186) and P53, relieving the repression of P53 and rescuing the proliferation and neuronal differentiation of NPCs. (B to E) Western blot analyses of p-MDM2 (B and C) and P53 (D and E) in Fmr1 WT and KO NPCs treated with Nutlin-3 (Nt3), showing that Nutlin-3 had no significant effect on p-MDM2 expression in Fmr1 KO and WT NPCs but specifically rescued P53 expression in Fmr1 KO NPCs without affecting WT cells (n = 3). GAPDH was used as a loading control in Western blot analyses. (F and G) Nutlin-3 treatment rescued the cell proliferation phenotype of Fmr1 KO NPCs, as demonstrated by immunostaining cells using the cell proliferation marker BrdU (F, red; scale bar, 20 μm), followed by quantitative analysis of BrdU+ cells (G) (n = 3). (H and I) Nutlin-3 treatment rescued neuronal differentiation phenotypes of Fmr1 KO NPCs, as assessed by a neuronal marker Tuj1+ (red; scale bar, 20 μm) (H); quantitative analysis in (I) (n = 3). (J and K) Nutlin-3 treatment specifically rescued astroglial differentiation phenotypes of Fmr1 KO NPCs, as assessed using the astroglial marker GFAP+ (green; scale bar, 20 μm) (J); quantitative analysis in (K) (n = 3). **P < 0.01; ***P < One-way ANOVA was used for all data analyses. Data are presented as means ± SEM.
Yue Li et al., Sci Transl Med 2016;8:336ra61
Published by AAAS