Figure 4.

CIN85 SH3 domains can bridge ASAP1 and ARAP3 ArfGAP proteins. (A) HEK293T cells were transiently transfected with GFP-ARAP3 or FLAG-ASAP1, wild-type or arginine mutants (WT or R, respectively), in the presence or absence of CIN85. Cell lysates were immunoprecipitated (IP) with anti-GFP antibodies, and after recovery of the bound material, detection was performed by immunoblotting (IB) with indicated antibodies. Expression of proteins was monitored in total cell lysates (TCL). (B) Experimental procedure was the same as in A, except that ΔCc or 3SH3-CIN85 constructs were used instead of the wild-type CIN85.