FIGURE 2.

Interaction of AGAP1 and AGAP2 with RhoA. A, a schematic of AGAP1 and AGAP2 is shown. The sequence identity for each domain is indicated. Ank, ankryrin repeat. B–D, shown is detection of RhoA-AGAP interaction by coimmunoprecipitation. In panel B, HeLa cells expressing AGAP1-HA and myc[T19N]RhoA were lysed. In panel i proteins were precipitated using an antibody to the myc epitope, and the relative quantities of AGAP1-HA in the precipitates (ppt) were determined by immunoblotting for the HA epitope. Blots for proteins in the total lysates are labeled IB HA and IB Myc. The blot for the HA-tagged AGAP1 that was precipitated with mycRhoA is labeled IP Myc and IB HA. In panel ii, proteins were precipitated with an antibody to the HA epitope, and the amount of RhoA in the precipitate was detected with an antibody to the myc epitope. In panel C, HeLa cells expressing AGAP2-GFP and myc[T19N]RhoA were examined. In panel i, proteins were precipitated with an antibody to GFP. In panel ii, proteins were precipitated with an antibody to myc. In panel D, cells expressing HA-AGAP2 (the epitope was fused to the N terminus of AGAP, as opposed to the epitope on the C terminus of AGAPs in other experiments presented in this figure) and myc[T19N]RhoA were examined. Proteins were precipitated with an antibody to the myc epitope. For all experiments, the input blots are 6% of the lysate used for the IP.