Figure 2.

Protein tyrosine nitration of the Ca²⁺ channel following in vitro treatment with peroxynitrite (ONOO-) in hCa_v1.2b-transfected CHO cells. (A) Immunoblot with anti-Ca_v1.2 antibody (left) and anti-nitrotyrosine antibody (right). Protein samples were immunoprecipitated with the Ca²⁺ channel antibody before immunoblot. Treatment with peroxynitrite enhanced tyrosine nitration of hCa_v1.2b. (B) Expression of phospho-CREB in hCa_v1.2b-transfected CHO cells detected by anti-pCREB antibody (1:50 dilution). Staining in the nucleus was observed in hCa_v1.2b transfected cells treated with 80 mM KCl for either 30 min or 60 min but not in the absence of depolarization (top panel). An Alexa Fluor 568 conjugated antibody (1:1000 dilutions) was used as the secondary antibody. Representative images from three experiments are shown. CREB, cyclic AMP response element binding protein.