Identification of PKCδ mutation and protein expression. (A) Sequencing of PRKCD with genomic DNA from blood from the index patient, parents, and 3 siblings. (B) Immunoblot analysis of PKCδ and of the control protein β-actin in the cells from the patient and normal volunteer. Purified B cells were activated with F(ab′)2 specific for IgM (10 μg/mL), IL-4 (50 ng/mL), and CD40 ligand (10 μg/mL) for 5 days, and then cell lysates were subjected to western blot (WB) analysis. The protein lysates from PBMCs and EBV-transformed B cells were used without stimulation. (C) PRKCD gene expression levels in the PBMCs and EBV-transformed B cells are expressed as a relative expression of normal cells. Data are represented as means ± SE of n = 3 separate experiments.
