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. Author manuscript; available in PMC: 2019 Sep 5.
Published in final edited form as: Sci Signal. 2018 Dec 18;11(561):eaat0358. doi: 10.1126/scisignal.aat0358

Fig. 2. Spontaneous local Ca2+ entry events via ORAI1.

Fig. 2.

(A) Non stimulated Jurkat Orai1-G-GECO1.2 T cells were imaged in 1 mM Ca2+ buffer (control), in nominal Ca2+ free buffer with 1 mM EGTA (no [Ca2+]ex), or in 1 mM Ca2+ buffer containing 100 μM Synta66 (Synta66). Furthermore, Jurkat Orai1-G-GECO1.2 T cells were loaded with 5 μM BAPTA-AM or incubated with 500 μM BZ194 overnight. Indicated time points in (A) represent dashed lines in (B). (B) F/F0 tracings of ROIs (5×5 pixel) from the five representative Jurkat Orai1-G-GECO1.2 T cells in (A). Arrowheads in (A) indicate the ROIs used in (B). Noise in the tracing was reduced by a moving average filter (width=5). (C) Statistical analysis of four ROIs per cell (20 × 10 pixel) over 300 s from control (n=31), no [Ca2+]ex (n=17), Synta66 (n=18), BAPTA (n=18) and ) and BZ194 (n=8). Statistically significant differences between the number of signals/ROI/s are marked by asterisks (* p < 0.05, ** p<0.01, *** p<0.001, Kruskal-Wallis Test). Length scale bar in (A) represents 10 μm.