Fig. 1
From: AAV-compatible optogenetic tools for activating endogenous calcium channels in vivo
Reducing the size of monSTIM1 by replacing GFP with a small tag. A Schematic of the working mechanism of GFP-monSTIM1. B Sizes of coding sequences of monSTIM1 and components of the AAV cassette. C Fluorescence images of cells co-expressing RGECO1 and monSTIM1 variants or OptoCRAC. Blue light was delivered for 2 min at 30-s intervals, and changes in intracellular Ca2+ levels were monitored by imaging RGECO1 (left, magenta images). Expression of monSTIM1 variants and OptoCRAC (right, green images). D Summary data showing changes in the normalized intensity of RGECO1 over time upon transient activation of monSTIM1 variants and OptoCRAC. E Summary data showing maximum fold changes (ΔF/F0) in RGECO1 intensity for monSTIM1 variants and OptoCRAC upon blue light stimulation (EGFP-monSTIM1: n = 18; FLAG-monSTIM1: n = 23; HA-monSTIM1: n = 24; OptoCRAC: n = 31; FLAG-monSTIM1(CRY2D387A): n = 38 cells). F, G Quantification of activation F and deactivation G kinetics of monSTIM1 variants (EGFP-monSTIM1: n = 30–38; FLAG-monSTIM1: n = 42–50; HA-monSTIM1: n = 45–47 cells). H Summary data showing the light-sensitivity of each optogenetic module. Data are presented as means ± SEM (*p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001; Student two-tailed t test); ns, not significant (p > 0.05)