(A) Dot blot analysis of lysates from HEK293 cells, separately transfected with the indicated individual γ-Pcdh-GFP fusion construct. N144/17 is monospecific for γ-Pcdh-A3. N144/32 detects 11 of the 12 proteins in the A subfamily, but none in the B or C subfamilies. N148/30 is monospecific forγ-Pcdh-B2. N159/5, raised against the constant exon, detects all γ-Pcdhs as expected, similar to a polyclonal antibody raised against the same antigen. GFP antibody detects all transfected cell lysates. (B–E) Western blots of whole brain lysates from either wild-type (WT) or Pcdh-γdel/del (KO) neonates. All of the monoclonal antibodies recognize appropriately sized bands in WT, but not KO, brain, confirming the their specificity. (F) Tubulin loading control blot. (G) Western blot of HEK293 cell lysates transfected with N-terminal HA-tagged full-length (FL) or truncated (ΔC) γ-Pcdh construct lacking the constant domain. N159/5 detects the full-length γ-Pcdh, but not the truncated construct, as expected.