Fig. 6. P2Y₂R-mediated G₁₂ activation requires v integrin expression and activity. (A) Cells expressing the WT P2Y₂R were incubated overnight at 37°C in serum-free medium with 10 µg/ml anti-v5 or anti-3 antibody, then stimulated with 100 µM UTP for 2 minutes. G₁₂ phosphorylation was detected as described in Fig. 5. Relative intensities are shown below each band as mean ± s.e.m. of results from four experiments. (B) Cells expressing the WT P2Y₂R were transfected with 5 µg of v antisense (AS) or sense (S) oligonucleotides, as described previously (Bagchi et al., 2005). Lipofectamine transfected cells served as a negative control. Expression of v integrin in these cells was determined by immunoblot analysis and a representative blot is shown. UTP-induced binding of [³⁵S]GTPS to G₁₂ was determined, as described in Fig. 5. Data are the mean ± s.e.m. of results from three separate experiments and are shown as fold increase over untreated cells.