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Fig. 2. Cdk5 is necessary for the differentiation of primary OLs. (A) After induction of differentiation, primary OL morphology was observed using phase-contrast microscopy. Representative images of cells in each category are shown. (B) Percentages of the cells treated with or without 10 µM roscovitine in each category after induction of differentiation. (C) Primary OLs treated with or without 10 µM roscovitine immunostained with anti-MBP antibody (red) before or 3 days after induction of differentiation. Cells can only be identified by DAPI staining of the nuclei (blue). (D) Percentages of MBP-positive cells (red) from the total number of cells. Data were evaluated using Student's t-test (*P<0.01). (E) After induction of differentiation, primary OLs treated with or without 10 µM roscovitine were lysed and immunoblotted with antibodies against MBP, PLP1 and β-actin. (F) Primary OPCs were infected with Cdk5, p35 or luciferase shRNA retroviral vectors. At 3 days after induction of differentiation, lysates from the infected cells were immunoblotted with antibodies against Cdk5, p35 or β-actin. (G) Representative fluorescence images of MBP- and GFP-positive cells are shown. Arrows indicate GFP-expressing cells. (H) Percentages of MBP-positive cells (red) against the total number of infected cells. Data were evaluated using Student's t-test (*P<0.01).
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