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. 2018 Sep 5;8:13252. doi: 10.1038/s41598-018-31621-0

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© The Author(s) 2018

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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Overexpression of the IP3R enhances STIM1 puncta formation. (A) Total internal reflection microscopy (TIRFM) images of STIM1-CFP forming puncta. Each panel shows a single cell. Scale shows 5 microns (μ). (B) Examples of individual puncta obtained from cells like those shown in (A). Each panel shows a single punctum. Middle panel shows the average of at least 120 individual puncta obtained from at least 15 independent cells. The panel to the right shows the 3D projection of the average puncta for each condition. The height shows the intensity in pseudo color scale (shown to the extreme right). The conditons explored are puncta formation induced by thapsigargin (TG), bradykinin (Bk) and Bk in cells overexpressing the IP3R. (C) Example of a single puncta, illustrating the co-localization of STIM1-CFP and YFP-IP3R. The right panel shows a pseudo color scale of the pixel co-localization according to Pearson coefficients (material and Methods).