Real time RT-PCR and western immunoblot of ganglia and cultures. A, real time RT-PCR of ganglia and cultures from rat and mouse. Ordinate: relative increment with respect to the ganglion products. For each receptor, amplification values were normalized with β-tubulinIII mRNA levels and compared with the ganglion mRNA levels. GAPDH amplification control was the same in all reactions. B, western immunoblots of equal amounts of neuronal protein lysates (β-tubulinIII) derived from ganglia or culture. Immunodetection of P2X3, P2X2 and TRPV1 mature proteins revealed proper migration (a) in accordance with Vulchanova et al. for the predominant P2X3 form (57 kDa), with Newbolt et al.  for the P2X2 mature receptor (65 kDa) and with Kedei et al.  for the TRPV1 mature receptor (120 kDa). Panel B b shows P2X3 native (45 kDa) and intermediate polypeptides (up to 50 kDa) detected one d in culture (1) but not in the ganglion (T) as reported by Nicke et al.  C, relative optical density values of mature receptors at 1–4 days in culture (normalized with respect to β-tubulinIII) and compared to ganglion values. For all experiments shown in A-C n = 3 animals for ganglion or day in culture datapoint (each point is mean ± SEM). *: P < 0.05; **: P < 0.01.