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. 2016 Dec 8;7:13725. doi: 10.1038/ncomms13725

Figure 2. The Orai1-ANSGA mutant is fully active and independent of STIM1.

Figure 2

(a) I–V relationship in store-replete HEK cells expressing CFP-Orai1-ANSGA. (b) I–V relationship in store-depleted HEK cells expressing CFP-Orai1-WT and STIM1-YFP. (c) Summary of the peak current amplitude at −100 mV. (d) Summary of the reversal potential measured from I–V data for Orai1-WT activated by store depletion and constitutively active Orai1-ANSGA in store-replete cells. (e,f) Fura-2 Ca2+ responses in stable HEK-STIM1-YFP cells expressing CFP-Orai1-WT (e) or CFP-Orai1-ANSGA (f) before and after store depletion with 2.5 μM ionomycin. (g) Summary of the constitutive (before ionomycin) and store-dependent (after ionomycin) Ca2+ entry in cells shown in e,f. (h) CFP fluorescence intensity of cells in e,f. (i,j) Internal BAPTA dialysis to deplete stores caused ICRAC to develop over time in HEK-STIM1-YFP cells expressing CFP-Orai1-WT. (k,l) With identical BAPTA-treated HEK-STIM1-YFP cells expressing CFP-Orai1-ANSGA, maximal ICRAC was observed at break-in. I–V curves (j,l) were determined at arrows shown (i,k). All values are means±s.e.m.