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. 2011 Mar 7;192(5):797–811. doi: 10.1083/jcb.201007014

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© 2011 Laursen et al.

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Figure 1. — Activation of β1-integrin enhances MBP expression and the morphological differentiation of oligodendrocytes. (A) Alignment of the transmembrane (blue) and intracellular domains of human β3- and β1-integrin. The mutated aspartic acid (D759) is shown in red. (B) Oligodendrocytes generated by OPCs cotransfected with EGFP cDNA plus an empty vector, wild-type, or mutated β1-integrin(D759R) as indicated. The cells were cultured on laminin and stained for EGFP and MBP. Note the increased process formation and generation of MBP+ myelin sheets in the cells expressing activated integrin. (C) Quantitative analysis of the morphology of oligodendrocytes generated by OPCs cotransfected with EGFP cDNA plus an empty vector, wild-type, or mutated β1-integrin(D759R or D759A). After transfection, OPCs were plated on PDL or laminin (Ln) for 3 d to allow differentiation. The morphology was scored as “simple,” “complex,” or “membrane” (Olsen and ffrench-Constant, 2005) based on their staining for EGFP and MBP. At least 100 EGFP+ cells were scored in each experiment. Results are an average of three independent experiments ±SD with statistical significance analyzed by two-way ANOVA followed by a Bonferroni post-test. *, P < 0.05; **, P < 0.01; ***, P < 0.001. Note that the D759R mutant enhances morphological differentiation in a ligand-independent manner. (D) Cells cotransfected as in C and analyzed for MBP expression. In each experiment at least 100 EGFP+ cells were scored and the percentage of MBP+ cells is given. The results are an average of three independent experiments ±SD with statistical significance tested by one-way ANOVA followed by a Tukey’s multiple comparison test. *, P < 0.05; **, P < 0.01; ***, P < 0.001. Note that both the D759A and D759R mutants enhance MBP expression in cells cultured in the absence of laminin. (E) Western blots for MBP of lysates from nontransfected OPCs differentiated for 1–4 d (D1–D4) in the absence or presence of laminin. Note the accelerated expression of MBP on laminin at D2. (F) MBP mRNA RT-PCR using mRNA from nontransfected OPCs differentiated for 0–3 d (D0–D3) in the absence or presence of laminin. Note that MBP mRNA levels are not affected by the laminin substrate.

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