L-ANAP–incorporating TRPV1-TAG-YFP channels retain functional properties. Cells were cotransfected with pANAP and a plasmid encoding TRPV1-Q169TAG-YFP, TRPV1-K240TAG-YFP, or TRPV1-S404TAG-YFP, as indicated, and were cultured with L-ANAP-ME in the medium. (A) Mean fluorescence data from cells transfected as described and loaded with the Ca2+ indicator Rhod-2-AM. 1 µM capsaicin (Caps.) was applied at 60 s, and ionomycin (Iono.) was applied at 240 s, as indicated by the arrows. Data represent means of 11, 6, and 10 cells for TRPV1-Q169TAG-YFP, TRPV1-K240TAG-YFP, and TRPV1-S404TAG-YFP, respectively. (B) Currents recorded in response to 1 µM capsaicin in inside-out excised patches held at 0 mV and jumped to potentials from −100 to 100 mV in steps of 20 mV. Currents in the absence of capsaicin were subtracted.
