Immunoblot analyses of CBFA2, pleckstrin, and PKC isozymes in platelets. (A) Immunoblot analysis of CBFA2 in platelets. Platelet lysates were subjected to SDS-PAGE and immunoblotting using an anti-CBFA2 antibody. Shown are the results in the patient (P) and 3 control subjects (N). (B) Immunoblot analysis of pleckstrin in platelets. Platelet lysates were subjected to SDS-PAGE and immunoblotting using an antipleckstrin antibody. Shown are the results in the patient (P), brother (B), sister (S), and 2 control subjects (N). 50% B and 50% N represent lanes with 50% protein applied. Pleckstrin levels are normal in the propositus. (C) Immunoblot analysis of PKC isozymes. The details are as described for panel B. PKC-θ levels were decreased in the patient (arrow). The results shown are representative of at least 3 separate experiments. To confirm that protein loading was comparable between the patient and the control subjects, the immunoblots used for PKC-θ were stripped and reblotted using antibodies against PKC-δ and PKC-ζ, which showed that the levels of these PKC in the patient were comparable to those of the other subjects (not shown).