Fig. 8. Cx431 colocalization with actin filaments and actin binding proteins. CNCs were double immunostained for Cx431 (green, A,D,G,J,M,P) and either vinculin (B), ezrin (H), IQGAP-1 (K), -actinin (N) or drebrin (Q) in red. F-actin (E) was stained using rhodamine phalloidin. In the merged images (C,F,I,L,O,R), regions of Cx431 colocalization with actin or these various actin-associated proteins can be observed. These regions of colocalization were mostly found at areas of cell-cell contact and along cell processes (see white arrowhead in insets). Boxed regions are magnified twofold in the bottom insets. Scale bar: 50 µm. (S) NIH3T3 cell extracts were immunoprecipitated with a Cx431 antibody and western immunoblotted with a vinculin antibody. A band of the size expected for vinculin was observed in the Cx431 immunoprecipitate and in the total lysate. (T) NIH3T3 cell extracts were immunoprecipitated with an IQGAP-1 antibody and western immunoblotted with a Cx431 antibody. A band of the size expected for Cx431 was observed in the IQGAP-1 immunoprecipitate and in the total lysate.