Fig. 3: Optogenetic release of BK-channels.
From: An optogenetic method for the controlled release of single molecules
a, Schematic of the FLAG-BK-YFP-PhoCl-TMEM construct. Following uncaging, BK-channels secreted to the membrane were detected using antibodies in the medium. b, TIRF and brightfield images of a CV-1 cell expressing FLAG-BK-YFP-PhoCl-TMEM with antibodies against FLAG before and after UV illumination. c, Quantification of the number of anti-FLAG antibodies bound to control (1.2 × 10−4 ± 9 × 10−5, n = 22) and to UV-illuminated cells (2.9 × 10−4 ± 1.8 × 10−4, n = 29), N = 5. Significance was tested using the two-tailed Mann–Whitney U-test (***P = 10−4). d, Quantification of the number of anti-FLAG antibodies bound to the same cells before (1 × 10−4 ± 7.3 × 10−5) and after uncaging (2.9 × 10−4 ± 1.9 × 10−4), N = 4, n = 19. Significance was tested using the two-tailed paired Wilcoxon signed-rank test (***P = 0.001). e, Representative current traces of BK-channel at varying voltage, with the protocol shown in red, in both control and UV-illuminated cells. f, Quantification of current density (pA/pF) at 120 mV in control (37 ± 20, n = 6) and UV-illuminated cells (380 ± 172, n = 8), N = 4. Significance was tested using a two-tailed Mann–Whitney U-test (**P = 0.002). Data presented as mean ± s.d. Center line in the box plot represents the median, the box represents 25–75% of the data, whiskers represent 1.5 × interquartile range and the square box among data points represents the mean. Scale bars, 10 µm.
