Figure 5.

Reducing PAK1 expression decreases pLIMK. A, B, Efficacy of PAK1 shRNA was quantified in C6 cells because of low transfection efficiency of primary rat oligodendrocytes. Thus, PAK1 and its downstream target pLIMK expression was analyzed on Western blot of C6 cells after shRNA transfection. *p < 0.05 (Dunnett's multiple comparisons test). C, rOPCs were grown in rOPC media with PDGF/FGF for 2 d. Cells were transfected at the last 4 h; then the media was replaced with differentiation media + T3, which was replaced every 2 d; and the cells were collected for analysis after 3 d. D, rat OPCs transfected with a scrambled control shRNA (shPAK1-control) or each of the three different PAK1 targets, then differentiated for 3 d. Oligodendrocytes were then collected and stained for GFP to show the impact of shRNA transfection, using O1/O4 (blue) as an oligodendrocyte membrane marker and PAK1 (red). Scale bar, 10 µm.