Fig. 3.

ENT-A011 induces neurogenesis on mouse adult hippocampal NSCs. A, A′ ENT-A011 increases proliferation in adult hippocampal NSCs. Representative fluorescence microscopy images of Immunostaining for BRDU and Nestin in adult hippocampal NSCs treated with BDNF or ENT-A011 for 24 h with or without Aβ42 (A). Quantification of BRDU fluorescence change with or without Aβ42 (A′). Quantification represents BRDU positive cells normalized against Nestin positive cells. (without Aβ42: N = 7–8, with Aβ42: N = 3), error bars represent S.E.M., Student’s t-test against Control; *p < 0.05, **p < 0.01, ***p < 0.001. B, B′. The compound rescues adult hippocampal NSCs from Aβ-induced cell death. Representative fluorescence microscopy images of Celltox cytotoxicity assay on hippocampal NSCs in the presence of Aβ42, treated with BDNF or ENT-A011, with or without the TrkB inhibitor ANA-12 for 24 h (B). Quantification of fluorescence change and MTT levels after BDNF or compound treatment, with or without ANA-12 N = 3–5. (B′). Quantification represents Celltox positive cells normalized against Hoechst positive cells (without ANA-12: N = 5–8, with ANA-12: N = 3)., error bars represent S.E.M., Student’s t-test against Control; *p < 0.05, **p < 0.01, ***p < 0.001. C, C′ ENT-A011 promotes differentiation of adult hippocampal NSCs. Representative fluorescence microscopy images of Immunostaining for Tuj1 and GFAP in adult hippocampal neural stem cells treated with BDNF or ENT-A011 for 10 days (C). Fold change in expression of TUJ1 (N = 6 for untreated and BDNF, N = 5 for ENT-A011), MAP2 (N = 5) and GFAP (N = 4) assessed by RT-qPCR in BDNF or ENT-A011 treated adult hippocampal NSCs relative to untreated control cells. (C′) Scalebars = 20 μm