Fig. 4: Optogenetic release of VRACs.
From: An optogenetic method for the controlled release of single molecules
a, Composition (left) and activation of VRACs (right). b, Schematic of VRAC release using PhoCl. VRACs comprising LRRC8A-PhoCl-TMEM and LRRC8E-GFP are expressed in LRRC8 knockout (KO) cells. Following uncaging, VRACs traffic to the plasma membrane (PM). c, Quantification of current density (pA/pF) at −80 mV following 50% hypotonic shock in untransfected HEK293 LRRC8 knockout cells (1 ± 0.5, n = 7, N = 1) and in cells transfected with LRRC8A-PhoCl-TMEM and LRRC8E-GFP, without (control) (1 ± 0.7, n = 12, N = 2) and with UV illumination (5.6 ± 4.9, n = 10, N = 3). Significance was tested using a two-tailed Mann–Whitney U-test (control versus UV, *P = 0.03, untransfected versus UV, *P = 0.04). d, Current traces of activated VRAC measured using the protocol shown in red in a control and a UV-illuminated cell. e, Time course of VRAC currents measured using voltage ramp protocol (shown in red) under isotonic condition, following 50% hypotonic shock and application of DCPIB, in a representative UV-illuminated cell. 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.
