Figure 2
Control of Ca2+ release with blue light and comparison with opto-α1AR.
(A) Subcellular localization of nMagHigh1-mKikGR-CAAX at the plasma membrane (upper panel) and its line profile (lower panel). (B–E) A comparison between the present approach based on the Magnet system and conventional opto-α1AR. For Ca2+ imaging, the cells were expressed with R-GECO1 and continuously exposed to excitation light at 559 nm. In the Magnet-based approach, the response of R-GECO1 was observed when the cells were stimulated with activation light at 473 nm (B) but not observed in the absence of the activation light (C); In opto-α1AR, the response of R-GECO1 was observed with (D) or without (E) blue light illumination at 473 nm, suggesting the excitation light of R-GECO1 at 559 nm activates opto-α1AR. The insets are partially enlarged views in Figure D and E. (F,G) Ca2+ imaging using the Magnet based approach with pulsed blue light illumination. Time-lapse fluorescence imaging showed that repeatable response of R-GECO1 was evoked upon activation light at 473 nm. Blue bars indicate 473 nm laser illumination.
