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. 2021 Jan 5;12:573714. doi: 10.3389/fnsyn.2020.573714

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Copyright © 2021 Kushnireva, Korkotian and Segal.

This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.

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Averaged STIM 1&2 fluorescence in protrusion, base, and shaft dendrites, in normal medium (2 mM Ca) and 15 min after incubation with a calcium-free medium. (A) STIM1 (red) and 2 (green) fluorescence in cell incubated with and without calcium, 10 DIV. (B) Bar graphs: averaged fluorescence minus background for in each group, 10 DIV. The difference between STIM1 and 2 in the base in both conditions and the difference between STIM2 with and without calcium in protrusion is highly significant (n = 10 dendrites from six cells for each group, ANOVA p < 0.001). (C) STIM1 (red) and 2 (green) fluorescence in a medium with and without calcium, 20 DIV. (D) Bar graphs: averaged fluorescence minus background for in each group, 20 DIV. The difference between STIM1 and 2 in the base in both conditions is not significant, but the difference between STIM2 with and without calcium in protrusion is highly significant (n = 8 dendrites from four cells for each group, ANOVA p < 0.001). (E) An example of the moving of STIM1/2 puncta in a protrusion (spine) in medium with and without calcium and after the return of calcium back to normal. White arrows mark protrusions in which the influx of STIM2 in a calcium-free medium is most noticeable (A,C, right panels). (F) STIM1 and STIM2 fluorescence at the base with protrusions (filopodia or spines) with background subtracted, at 10 DIV, 15, and 20 DIV. DIV 10: 34 protrusions; DIV 15: 30; DIV 20: 35 protrusions. (G) An example fluorescence calculation of STIM1/2 puncta at the base with protrusions (spine) for (F). *Significant, 0.05 > p > 0.01; ***highly significant, p < 0.001; n.s., not significant, p ≥ 0.05.