Fig. 4.

PCDH-α, PCDH-β, and PCDH-γ isoforms interact with each other in vitro. A, co-IP experiments show that PCDH-α isoform C2, PCDH-γ isoform C5, and PCDH-β isoform 22 interact with each other in transfected cells. HEK293T cells were co-transfected with different combinations of expression vectors followed by co-IP using the anti-FLAG M2 affinity resin and Western blot with anti-V5, anti-pan-PCDH-α, and anti-pan-PCDH-γ antibodies. B, interactions among different PCDH isoforms are calcium-independent. Similar experiments as in A were carried out with an addition of 20 mm EGTA in the lysis buffer to deplete calcium.