FIG 3.

CRB3A recruits the Ehm2/p114RhoGEF signaling module to cell-cell contacts. (A) Flag-Ehm2 was immunoprecipitated (IP) from control or CRB3A-expressing HeLa cells. Western blotting detected immunopurified Flag-Ehm2, as well as p114RhoGEF and CRB3A that were coimmunoprecipitated. (B to I) Immunofluorescence of Ehm2 or p114RhoGEF in control HeLa cells (empty vector) or HeLa cells expressing Myc-CRB3A, Myc-CRB3A^ΔERLI, or Myc-CRB3A^MutFBM. (J) Western blot analysis validating Ehm2 or p114RhoGEF knockdown in Myc-CRB3A-expressing HeLa cells. Actin was used as a loading control, whereas Western blotting using an anti-Myc antibody shows that Myc-CRB3A expression is maintained in knocked down cells. (K and L) Immunofluorescence of p114RhoGEF in Myc-CRB3A-expressing HeLa cells transfected with a scrambled siRNA (control RNAi) or knockdown for Ehm2 (Ehm2 RNAi). (M and N) Immunostaining of Ehm2 in Myc-CRB3A-expressing HeLa cells knocked down for p114RhoGEF (p114RhoGEF RNAi). A scrambled siRNA was used as a control (control RNAi). Scale bar, 20 μm.