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. 2019 Jul 19;10:3223. doi: 10.1038/s41467-019-11207-8

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© The Author(s) 2019

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.

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Fig. 2 — Orai2 is not required for RyR-dependent Ca²⁺ release from internal stores. a Left: Caffeine was locally applied to the somata of whole-cell patch-clamped CA1 pyramidal neurons (PNs) filled with OGB-1 through the patch pipette. Black traces: Fluorescence transients in response to three consecutive caffeine applications with an interval of 2 min in a wild-type mouse. Red traces: Analogous experiment in an Orai2-deficient mouse. b Summary of the experiments in a. Bar graph shows the mean amplitudes of relative fluorescence changes resulting from the first caffeine application in each cell (n = 20 cells in wild-type and n = 28 cells in Orai2−/− mice, p = 0.268). c Left: Both caffeine and ultraviolet pulses were applied to somata of whole-cell patch-clamped CA1 PNs filled through the patch pipette with NPE (caged)-IP₃ and OGB-1. Red traces: Fluorescence transients resulting from three cycles of alternating somatic IP₃ uncaging and caffeine applications in an Orai2−/− mouse. d Summary of experiments in c. Bar graph showing the mean amplitudes of responses to caffeine (white) and IP₃ uncaging (purple; n = 18 cells). e Left: Both caffeine and 3,5-dihydroxyphenylglycine (DHPG) were locally applied to the somata of whole-cell patch-clamped CA1 PNs filled with OGB-1 through the patch pipette. Traces: Fluorescence transients in response to somatic caffeine and DHPG applications with the indicated time intervals in control artificial cerebrospinal fluid (left traces) and in the presence of 10 µM ryanodine (right traces). f Summary bar graph showing mean amplitudes of relative fluorescence changes for all experiments (n = 10 cells, p = 7.89 × 10⁻⁹) in e. *** - extremely significant (p < 0.0001)