Colocalization of mSlo1 and TRPV1 in HEK293 cells.

<p>A, Surface expression of mSlo1 and TRPV1. HEK293 cells were cotransfected with mSlo1-TagGFP (green) and TRPV1-myc (red). Confocal images of mSlo1-TagGFP (left panel) and TRPV1-myc (middle panel) have clearly green and red edges, respectively, and the merged image (right panel) shows a yellow edge, indicating that two proteins were colocalized. 132 colocalized cells from 3 experiments were imaged and analyzed. Scale bar is 5 μm. B, coimmunoprecipitation experiments carried out on lysates of TRPV1 and mSlo1 co-expressed HEK293 cells. Left: TRPV1 channels can be detected in immunoprecipitates prepared by anti-myc antibody (mSlo1-IP) (n = 4). Right: mSlo1 channels can be detected in immunoprecipitates prepared by anti-HA antibody (TRPV1-IP) (n = 5). The lane marked “input” refers to a sample of the original cell lysate from expressed HEK293 cells. The lane “IgG-IP” respresents a negative control sample, which was immunoprecipitated with rat serum not expressing epitope-tagged proteins. The lane “mSlo1”, “TRPV1” and “mock”respresent samples of transfected alone or no transfected HEK293 cells. Note that no signal was detected from IgG-IP or mock samples. C, The currents of TRPV1 were not affected by myc. The representative trace of TRPV1-myc was obtained at -60 mV, in the presence of 5 μM Capsaicin. D, The currents of TRPV1/mSlo1 were not affected by HA. The representative traces of TRPV1-HA/ Slo1-myc were obtained by a voltage step from -60 to 0 mV, in the absence and presence of 5 μM Capsaicin. (n=4-8).