Figure 5.

Tyr-410 phosphorylation of p130Cas is specific to Gα_i2. HeyA8 (2 × 10⁶) cells were transiently transfected with either a vector control or (A) Gai2Q205L, (B) Gα_qQ209L, (C) Gα₁₃Q226L, or (D) Gα₁₂Q229L and incubated in 6-well plates in normal growth media. After approximately 48 hours, the cells were lysed and subjected to immunoblot analysis using antibodies specific to Tyr-410–phosphorylated p130Cas and then stripped and probed for total p130Cas. Overexpression of each constitutively active G protein was confirmed with antibodies to Gai2, Gα_q, Gα₁₂, or Gα₁₃ on their respective blots. The blots were stripped and reprobed with antibodies to GAPDH to monitor equal loading of proteins. (E) Quantification and statistical analysis of the experiments represented in A to D (n = 3).