TRPC6 expression in human neutrophils. (A) Expression of TRP family members in isolated human neutrophils. Representative results of RT-PCR analysis with mRNA from a single isolation and cDNA preparation of neutrophils with each of the indicated primers are given and GAPDH was used as a positive control. Lanes are indicated as: M, markers; 1, TRPC1; 2, TRPC2; 3, TRPC3; 4, TRPC4; 5, TRPC5; and 6, TRPC6. (B) Western blotting of neutrophil (N) or mononuclear cell (M) membrane preparations stained with anti-TRPC6 antibody (1:400) revealed a strong band at the predicted molecular weight (100 kDa) as indicated. Specificity was demonstrated by incubation of the TRPC6 antibody with a 4-fold excess of the antigenic peptide exhibited no signal. Nonspecific binding was assessed using an IgG control stained with rabbit anti-IgG antibody (1:400). A representative immunoblot of 3 different experiments is shown.