Fig. 7: ORAI2 and ORAI3 ensure differential activation of NFAT isoforms.
From: The native ORAI channel trio underlies the diversity of Ca2+ signaling events
a, d Time-lapse quantification represented as mean ± SEM of nuclear translocation in response to 10 µM Cch stimulation in the presence of 2 mM Ca2+ of NFAT1-GFP transfected either in parental HEK293 cells or ORAI single- (a), double- (d), or triple- (d) knockout cells; images were captured every 10 s. b, e Maximal NFAT1-GFP nuclear translocation determined by the ratio of nuclear/total cell GFP fluorescence. c, f Rate of NFAT1-GFP nuclear translocation in response to Cch stimulation. g Representative images of different cell types transfected with NFAT1-GFP at time 0 and 15 min and 30 min after stimulation with 10 µM Cch. Scale bar: 10 µm (for all NFAT1 experiments n = 98(HEK293), n = 84(ORAI1-SKO), n = 93(ORAI2-SKO), n = 44(ORAI3-SKO), n = 79(ORAI1,2-DKO), n = 65(ORAI1,3-DKO), n = 76(ORAI2,3-DKO), n = 64(ORAI-TKO); all points are single cells). h, k Time-lapse quantification represented as mean ± SEM of nuclear translocation in response to 10 µM Cch stimulation in the presence of 2 mM Ca2+ of NFAT4-GFP transfected either in parental HEK293 cells or ORAI single- (h), double- (k), or triple- (k) knockout cells; images were captured every 10 s. i, l Maximal NFAT4-GFP nuclear translocation determined by the ratio of nuclear/total cell GFP fluorescence. j, m Rate of NFAT4-GFP nuclear translocation in response to Cch stimulation. n Representative images of different cell types transfected with NFAT4-GFP at time 0, and 15 min and 30 min after stimulation with 10 µM Cch. Scale bar: 10 µm (for all NFAT4 experiments n = 130(HEK293), n = 72(ORAI1-SKO), n = 82(ORAI2-SKO), n = 79(ORAI3-SKO), n = 90(ORAI1,2-DKO), n = 59(ORAI1,3-DKO), n = 71(ORAI2,3-DKO), n = 59(ORAI-TKO); all points are single cells). Scatter plots are represented as mean ± SEM. All data in Fig. 7 was analyzed using the Kruskal–Wallis one-way ANOVA with multiple comparisons and was derived from a minimum of three independent experiments. (*p < 0.05; **p < 0.01; ***p < 0.001; ****p < 0.0001).
