Western blot detections of TRPV1 protein in undifferentiated (-) and differentiated (diff) PC-12 cells (A), wild-type (WT) and TRPV1 KO (G12, H3) A10 cells (A and B), in growing (gr) and quiescent (quiesc) rat primary VSMCs (A and B), and in TRPV1 transfected HEK293T cells (B) using two different antibodies against TRPV1: mouse monoclonal antibody (mAb) against the C-terminus (A, upper panel) and rabbit polyclonal antibody (pAb) against N-terminus (A, lower panel and B), α/β-tubulin was used as a loading control. In each experiments cells of different passages were used. The membrane in experiment 3 was cut in the attempt to reduce unspecific binding by exposing only the part where TRPV1 was expected to bind the primary antibody. (b) One representative blot out of 4 independent western blot experiments with similar results is shown. (C) Capsaicin concentration-response curves measured by calcium microfluorimetry. The response to capsaicin was quantified by the area under the curve (AUC) and fitted by a sigmoidal function. HEK293T cells transfected with human TRPV1 (red) respond to capsaicin, in contrast to untransfected HEK293T cells (black).