Fluorescent TRPV1 channels in unroofed cells. (A) Epifluorescent images of the same field of unroofed cells transfected with TRPV1-Q169TAG-YFP and pANAP and cultured with L-ANAP-ME in the medium in the ANAP color channel (left) and the YFP color channel (right). The top images show the initial data collected for this field. The bottom images show the field after a 10-s prebleaching UV exposure. The scale bar applies to all images and is 10 µm. (B) Collected data representing the dependence of fluorescence intensity in the ANAP color channel on cumulative time exposed to ANAP excitation light. Blue points represent YFP-negative cells, pink points represent TRPV1-K240TAG-YFP–expressing cells, and green points represent TRPV1-S404TAG-YFP–expressing cells. Solid curves are fits with a single exponential with a time constants of 10 s (n = 5 cells), 100 s (n = 3 cells), and 110 s (n = 6 cells) for YFP negative, TRPV1-K240TAG-YFP, and TRPV1-S404TAG-YFP, respectively. Error bars represent the SEM.