Fig. 1

Novel intracellular association of Pcdhs. a Indicated “test” constructs derived from γA3 were cotransfected with the γB2 full-length “target”, complexes were immunoprecipitated (IP) with anti-GFP beads, and blots were probed with the indicated antibodies. In the absence of γA3-GFP, γB2-RFP was not precipitated with anti-GFP beads, even though γB2-RFP was present in the total lysate (lane 1). γA3 full-length, constant domain deleted (Δconst), and the larger cytoplasmic deletion (Δ190) of γA3 all coprecipitated with full-length γB2 (lanes 2–4) confirming an extracellular interaction described previously [9]. However, the γA3 extracellular deletion (ΔECD, lane 5) as well as a “stub” construct consisting mostly of the variable cytoplasmic domain (VCDst, lane 6) also coprecipitated with γB2 indicating a novel cytoplasmic association. b Indicated γA3 test constructs derived were coprecipitated with the γA3 VCD stub. The stub associated with full length and constant domain deleted γA3 (lanes 2 and 3) but not with γA3Δ190 (lane 4). c The γA3 VCD stub was truncated at the indicated points (arrowheads) and tested for coprecipitation with full-length γA3 or γB2-RFP. Boxed region indicates the previously mapped trafficking motif (VCD motif). Truncation into the VCD motif eliminated coprecipitation with full length γA3 and γB2