Neutrophil PLC-β2 and PLC-γ2 levels.

(A) PLC-β2 mRNA levels in neutrophils. First-strand cDNA from neutrophils from the patient (P) and healthy subjects (H) was amplified by PCR using primers for PLC-β2 corresponding to 2864 nt to 2882 nt (forward) and 3328 nt to 3352 nt (reverse) along with primers for β-actin (758-bp fragment). The products were analyzed as described in the Figure 2 legend. P1 and P2 represent patient samples from 2 separate visits. Results shown are representative of 3 separate experiments. (B) Immunoblots showing PLC-β2 and PLC-γ2 levels in neutrophil lysates from patient (P) and 3 healthy subjects. The neutrophil lysates were subjected to electrophoresis and transferred to membrane. The PLC-β2 and PLC-γ2 were detected using specific antibodies and by chemiluminescence. Samples from healthy subject H1 were analyzed at increasing protein concentration from 0.5 μg to 12.0 μg subjected to gel electrophoresis. Samples from patient (P) and healthy subjects H2 and H3 were analyzed at 6 μg protein. Results shown are representative of 3 separate experiments.