Fig. 3

Chd7 regulates mouse NSPCs proliferation and self-renewal in vitro. a, Chd7 targeting using lentiviral shRNA significantly reduced NSPC growth compared to control shRNA, as assessed using a Cell Titer-Glo Assay Kit 4 days after infection. b, In the primary neurosphere formation assay, single dissociated cells of neurospheres generated from E14.5 brain were plated onto a 96-well plate and infected with lentiviruses encoding control shRNA or Chd7 shRNA. The cells were cultured in the presence of both EGF and FGF2 c, In the secondary neurosphere assay, neurospheres infected with a lentivirus encoding control shRNA or Chd7 shRNA were cultured in the presence of both EGF and FGF2 for 5 days. Then neurospheres were dissociated into single cells and seeded onto a 96-well plate at a cell density of 20 cells/μl in the presence of EGF and FGF2. d, Neurospheres infected with lentivirus expressing either control or Chd7 targeting shRNA were cultured in the presence of both EGF and FGF2 for 5 days and then dissociated and cultured onto the poly-D-lysine coated glass slip for 4DIV in the absence of growth factors. The gene expression of neural marker (NSE), an astrocyte marker (GFAP), or an oligodendrocyte marker (CNPase) was quantified by qRT-PCR. For the graphs, the data were compiled from three independent experiments. Error bars indicate S.E. values; **P < 0.01 versus control; Student’s t-test