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. Author manuscript; available in PMC: 2011 Oct 1.
Published in final edited form as: Arterioscler Thromb Vasc Biol. 2010 Aug 5;30(10):1976–1982. doi: 10.1161/ATVBAHA.110.212415

Figure 2. Role of PKC and Src in PDBU-induced GIT1 phosphorylation.

Figure 2

PDBU-induced GIT1 tyrosine phosphorylation in A7r5 (A) and in HEK293 (B) cells. Flag-GIT1 (WT) was transfected into A7r5 or HEK293 cells for 24 hours. The cells were starved for 6 hours prior to being treated with or without 1μM PDBU for various times. (C) Effect of PKC and Src inhibition on PDBU-induced GIT1 tyrosine phosphorylation. After transfection with Flag-GIT1 (WT), HEK293 cells were pretreated with 10 μM PP2 (Src inhibitor) or 5 μM Go6850 (PKC inhibitor) for 30 min and then treated with or without 1μM PDBU for 10 min. (D) Role of Src in PDBU-induced GIT1 tyrosine phosphorylation. SYF+/+ and SYF-/- fibroblasts were infected with Flag-GIT1 (WT) adenovirus for 24 hours and then starved for 6 hours followed by treatment with or without 1μM PDBU for 10 min. Lysates were immunoprecipitated with GIT1 antibody and then immunoblotted with 4G10 antibody to detect pGIT1 (top panels). To confirm equal protein immunoprecipitation, the blot was reprobed with GIT1 antibody (lower panels). The blots were analyzed by densitometry using LiCor software. Fold changes normalized to 0 min (A-B) or to the first lane 29 as shown below each blot (n=2-3).