Involvement of Sox10 and/or Nkx2.2 in the regulatory pathway of oligodendrocyte differentiation by DISC1.

A, B, Expression of Sox10 and Nkx2.2 mRNA were decreased by DISC1 overexpression. Oligodendrocyte precursor cells were infected with GFP-Adv or DISC1-GFP-Adv for 12 hours and induced to differentiate by depriving PDGF for 36 hours. C, D, Expression of Sox10 and Nkx2.2 mRNA were increased by DISC1 knockdown. Oligodendrocyte precursor cells were transfected with control-siRNA, DISC1-siRNA-1 or DISC1-siRNA-2 and cultured in medium containing PDGF for 48 hours. E, F, Expression of Sox10 and Nkx2.2 mRNA were increased by truncated DISC1 overexpression. Oligodendrocyte precursor cells were infected with GFP-Adv or trDISC1-GFP-Adv for 12 hours and induced to differentiate by PDGF deprivation for 36 hours. G–J DISC1 knockdown mediated increase of CNPase mRNA was inhibited by a simultaneous knockdown of either Sox10 or Nkx2.2. Oligodendrocyte precursor cells were co-transfected with control-siRNA or DISC1-siRNA-1 and sox10-siRNA or nkx2.2-siRNA and cultured in medium containing PDGF for 24 (H) or 48 hours (G, I, J). mRNA quantification was performed 48 hours after adenovirus infection (A, B, E, F) or siRNA transfection (C, D, G, I, J) or 24 h hours after siRNA transfection (H) by qRT-PCR. Data are expressed as mean ± s.e.m. of at least three independent experiments. *p<0.05 vs. GFP-Adv (A, B), *p<0.05 vs. control-siRNA (C, D), *p<0.05 (E–H).