Fig. 4: A fused Calcium sensor shows OLPVR1-delimited Ca2+ influx from the ER.
From: Hijacking of internal calcium dynamics by intracellularly residing viral rhodopsins
a Schematic representation of the calcium-imaging conditions tested. HEK293T cells were transfected with plasmids encoding for wild-type OLPVR1 and GCaMP6s (1&2), the fusion construct OLPVR1-GCaMP6s (3&4) or loss-of-function OLPVR1(K204Q)-GCaMP6s (5) and imaged with an epifluorescence microscope under 470-nm light in the absence (1,3&5) or presence (2&5) of 15 µM BAPTA-AM. b Time courses of fluorescence changes upon light application in the conditions described in a. c Confocal images of cells transfected with a plasmid coding for OLPVR1-GCaMP6s after 2 h incubation in 15 µM BAPTA-AM at T = 0, 15 and 30 s after light application. T = 0 is the first frame acquired after light is turned on. d Average maximal fluorescence changes upon light application of cells from at least 3 independent transfections in the conditions shown in a. Numbers of cells are in parentheses. (Error bars, SEM) e Half times of rise in fluorescence during illumination for conditions 1, 3 and 4. Numbers of cells are in parentheses (Error bars, SEM). **** p < 0.0001, one-way ANOVA, Tukey’s multiple comparisons test, FD (DFn 4, DFd 250) = 71.2; FE (DFn 2, DFd 143) = 68.8. Source data are provided as a Source Data file.
