Fig 2: Chemical constructions of the three compounds, NSCs culture and AD cell model establishment. Chemical constructions of the three compounds. (A) 3,6'-disinapoyl sucrose (DISS); (B) onjisaponin B (OB); (C) tenuifolin (TEN). Hippocampal cells were isolated from newborn C57BL/6 mice; and cultured in proliferation medium for 7 days to obtain neurospheres; and then dissociated the neurospheres into single cells cultured in differentiation medium for 7 days to test the multi-directional differentiation ability. To establish AD cell model; the neurospheres were transfected with APP gene; and RT-PCR and Western blot analysis were used to determine the transfection. (D) The NSCs were double positive for Sox-2 (green) and Nestin (red); Scale bar = 100 µm; (E) The neurons; astrocytes and oligodendrocytes were marketed by NF-M; GFAP and NG2; respectively. Scale bar = 25 µm. (F) The 293 T cells and NSCs transfected with APP-GFP and GFP plasmids expressed green fluorescent protein. Scale bar = 100 µm; (G) The Aß protein and mRNA levels in GFP-NSCs and APP-NSCs were measured by RT-PCR and Western blot assay; respectively; (H-I) Semi-quantification of Aß protein (H) and mRNA (I) as in G. **P < 0.01 vs. GFP-NSCs group. Data are represented as mean ± sem. Data were analyzed by t-test