Figure 1

Pten deletion promotes differentiation of NSCs into neurons and enlarges neurosphere. (A) Neurospheres generated from Pten ^loxp/loxp NSCs, were transfected with lentivirus expressing GFP (control) or Cre-GFP (for Pten deletion), and induced to differentiate by growth factor withdrawal. PTEN protein expression was abolished by Cre-GFP-mediated Pten removal, as detected by western blotting. GAPDH was used as a loading control. (B) The fate change of differentiated NSCs was ascertained by labeling with an antibody against the neuronal marker Tuj1. GFP⁺ cells represent the cells transfected with lentivirus. Nuclei were stained with DAPI. Scale bar = 40 μm. (C) A greater number of Tuj1⁺GFP⁺ cells was observed upon Pten deletion compared to Pten ^loxP/loxP controls. Data are expressed as mean ± SD. **P < 0.01 (n > 800 cells per group from each experiment). (D) Neurospheres cultured 3 days after harvesting through flow cytometry in proliferation condition culture. Scale bar = 100 μm. (E) Values are expressed as the neurospheres diameter. Data are expressed as mean ± SD. *P < 0.05 (The average sizes with standard deviations determined for all neurospheres in one well).