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. 2013 Apr;27(4):1532–1548. doi: 10.1096/fj.12-219600

Figure 5.

Figure 5.

MYLK is required for activation of Src and Pyk2. A, B) Src (A) and Pyk2 (B) phosphorylation in microglia isolated from WT or MYLK−/− mice. C) Tat-mediated increase in microglial adhesion was ameliorated in primary rat microglia pretreated with PP2 (1 μM), but not PP3 (1 μM). D) Right panel: effect of Tat on the cell adhesion of BV-2 cells in the presence of Src siRNA. Left panel: Western blot analysis of whole-cell lysates from BV-2 cells transfected with siRNAs Src or nonsense (non) siRNA using antibodies specific for Src. E) Effect of Tat on the cell migration of BV-2 cells in the presence of Src siRNA. F) Right panel: effect of Tat on the cell adhesion of BV-2 cells in the presence of Pyk2 siRNA. Left panel: Western blot analysis of whole cell lysates from BV-2 cells transfected with siRNAs Pyk2 or nonsense siRNA using antibodies specific for Pyk2. G) Effect of Tat on the cell migration of BV-2 cells in the presence of Pyk2 siRNA. H) Association of MYLK with Pyk2 or Src in primary rat microglia stimulated with Tat. I) Association of Pyk2 with MYLK or Src in primary rat microglia stimulated with Tat. Lysates were immunoprecipitated using anti-MYLK/Pyk2 antibody or followed by immunoblotting with anti Pyk2/Src/MYLK or IgG antibodies. Data are presented as means ± se of 4 independent experiments. *P < 0.05, **P < 0.01, ***P < 0.001 vs. control group; ###P < 0.001 vs. Tat-treated group.