KEY RESOURCES TABLE
| REAGENT or RESOURCE | SOURCE | IDENTIFIER |
|---|---|---|
| Antibodies | ||
| Mouse monoclonal anti-GAPDH | MilliporeSigma | Cat# MAB374; RRID: AB_2107445 |
| Rabbit monoclonal anti-STIM1 | Cell Signaling Technologies | Cat# 5668; RRID: AB_10828699 |
| Rabbit polyclonal anti-STIM2 | Cell Signaling Technologies | Cat# 4917; RRID: AB_2198021 |
| α-IP3R1 | Yule Lab | N/A |
| α-IP3R2 | Yule Lab | N/A |
| Anti-IP3R-3 | Becton, Dickinson (BD) | Cat# 610313; RRID: AB_397705 |
| α-Tubulin (DM1A) Mouse mAb | Cell Signaling Technologies | Cat# 3873S; RRID: AB_1904178 |
| 680 CW goat anti-Mouse | LI-COR | Cat# 926–68070; RRID: AB_10956588 |
| 800 CW donkey anti-Rabbit | LI-COR | Cat# 926–32213; RRID: AB_621848 |
| Chemicals, peptides, and recombinant proteins | ||
| Thapsigargin | Calbiochem | Cat#586005 |
| Carbachol | MilliporeSigma | Cat#212385 |
| Fura-2 AM | Invitrogen | Cat#F1201 |
| BAPTA-tetracesium Salt | Santa Cruz Biotechnology | Cat#480436-84-8 |
| Intercept® (TBS) Blocking Buffer | LI-COR | Cat#927–60001 |
| Halt Protease and Phosphatase Inhibitor | Thermo Scientific | Cat#PI78443 |
| RIPA Buffer | Sigma-Aldrich | Cat#R0278–50ML |
| Gadolinium(III) Chloride | ACROS Organics | Cat#AC383560050 |
| Cyclopiazonic Acid | Alomone Labs | Cat#C-750 |
| Lipofectamine 2000 Transfection Reagent | Thermo Scientific | Cat#11668027 |
| Cal-520, potassium salt | AAT Bioquest | Cat#21140 |
| ci-IP3/PM | Tocris | Cat#6210 |
| EGTA, AM | Invitrogen | Cat#E1219 |
| Critical commercial assays | ||
| Guide-it Mutation Detection Kit | Clontech Laboratories | Cat#631443 |
| Cell line Nucleofector Kit | Lonza | Cat#VCA-1003 |
| In-Fusion® HD EcoDry Cloning System | Clontech Laboratories | Cat#639688 |
| Pierce Rapid Gold BCA Protein Assay Kit | Thermo Scientific | Cat#A53225 |
| Deposited data | ||
| http://data.mendeley.com/login?redirectPath=/datasets/9dtvhhkzdz/draft?a=c43ec1d3-e142-4fd0-b29f-86660a9ecbe4 | Trebak Lab, this paper | N/A |
| Experimental models: cell lines | ||
| HEK293 cells | ATCC | Cat#CRL-1573 |
| STIM1 KO cells | Trebak Lab (Emrich et al., 2019) | N/A |
| STIM2 KO cells | Trebak Lab (Emrich et al., 2019) | N/A |
| STIM1/2 KO cells | Trebak Lab (Emrich et al., 2019) | N/A |
| Orai2,3 DKO cells | Trebak Lab (Yoast et al., 2020b) | N/A |
| Orai1,3 DKO cells | Trebak Lab (Yoast et al., 2020b) | N/A |
| Orai1,2 DKO cells | Trebak Lab (Yoast et al., 2020b) | N/A |
| Orai TKO cells | Trebak Lab (Yoast et al., 2020b) | N/A |
| Orai2,3-STIM1 KO cells | Trebak Lab, this paper | N/A |
| Orai2,3-STIM2 KO cells | Trebak Lab, this paper | N/A |
| Orai1,3-STIM1 KO cells | Trebak Lab, this paper | N/A |
| Orai1,3-STIM2 KO cells | Trebak Lab, this paper | N/A |
| Orai1,2-STIM1 KO cells | Trebak Lab, this paper | N/A |
| Orai1,2-STIM2 KO cells | Trebak Lab, this paper | N/A |
| IP3R TKO cells | Yule Lab(Alzayady et al., 2016) | N/A |
| Oligonucleotides | ||
| Forward primer for CMV-YFP-STIM2.1 (D80A/D82A) 5′-ACAAATGGCTGATGCCAAAGATGGTGGAATTG-3′ | Trebak Lab, this paper | N/A |
| Reverse Primer for CMV-YFP-STIM2.1 (D80A/D82A) 5′-TGGCATCAGCCATTTGTTTATGTATTGTTTGAAG-3′ | Trebak Lab, this paper | N/A |
| Forward primer for CMV-STIM1-F394H (D76A/D78A)-YFP 5′-ACTGATGGCCGATGCTGCCAATGGTGATGTGG-3′ | Trebak Lab, this paper | N/A |
| Reverse primer for CMV-STIM1-F394H (D76A/D78A)-YFP 5′-CAGCATCGGCCATCAGTTTGTGGATGTTACGG-3′ | Trebak Lab, this paper | N/A |
| ORAI1 g1 CRISPR/Cas9 Guide RNA 5′-GTTGCTCACCGCCTCGATGT-3′ | Trebak Lab, this paper | N/A |
| ORAI2 g1 CRISPR/Cas9 Guide RNA 5′-ACGACAGGGCCTGTACCGAG-3′ | Trebak Lab, this paper | N/A |
| ORAI2 g3 CRISPR/Cas9 Guide RNA 5′-CTCATGCGGGGACTCGCTGA-3′ | Trebak Lab, this paper | N/A |
| ORAI3 g2 CRISPR/Cas9 Guide RNA 5′-GTTCGTGCACCGCGGCTACC-3′ | Trebak Lab, this paper | N/A |
| ORAI3 g4 CRISPR/Cas9 Guide RNA 5′-CCAGAGACTGCACCGCTACG-3′ | Trebak Lab, this paper | N/A |
| STIM1 g3 CRISPR/Cas9 Guide RNA 5′-TGATGAGCTTATCCTCACCA-3′ | Trebak Lab, this paper | N/A |
| STIM2 g1 CRISPR/Cas9 Guide RNA 5′-AGATGGTGGAATTGAAGTAG-3′ | Trebak Lab, this paper | N/A |
| Recombinant DNA | ||
| HA-NFAT1(4–460)-GFP | Addgene | Cat#11107 |
| HA-NFAT4(3–407)-GFP | Addgene | Cat#21664 |
| LentiCRISPR v2 | Addgene | Cat#52961 |
| pU6-(Bbs1)-mCherry | Addgene | Cat#64324 |
| pSpCas9(BB)-2A-GFP (PX458) | Addgene | Cat#48138 |
| pCMV R-CEPIA1er | Addgene | Cat#58216 |
| CMV-CFP-ORAI1 | Gill Lab | NA |
| CMV-CFP-ORAI2 | Gill Lab | NA |
| CMV-CFP-ORAI3 | Gill Lab | NA |
| CMV-STIM1-YFP | Gill Lab | NA |
| CMV-STIM2-YFP | Gill Lab | NA |
| CMV-YFP-S1N-S2C | Trebak Lab(Emrich et al., 2019) | NA |
| CMV-YFP-S2N-S1C | Trebak Lab(Emrich et al., 2019) | NA |
| TK-YFP-S1N-S2C | Trebak Lab, this paper | NA |
| TK-YFP-S2N-S1C | Trebak Lab, this paper | NA |
| TK-STIM1-YFP | Trebak Lab, this paper | NA |
| TK-STIM2-YFP | Trebak Lab, this paper | NA |
| CMV-YFP-STIM2.1 | Gill Lab | NA |
| CMV-STIM1-F394H-YFP | Gill Lab | NA |
| CMV-YFP-STIM2.1 (D80A/D82A) | Trebak Lab, this paper | NA |
| CMV-STIM1-F394H (D76A/D78A)-YFP | Trebak Lab, this paper | NA |
| Software and algorithms | ||
| Graphpad Prism 8 | GraphPad | https://www.graphpad.com/scientific-software/prism/ |
| Clampfit 10.3 | Molecular Devices, LLC. | https://www.moleculardevices.com/ |
| ImageJ | (Schneider et al., 2012) | https://imagej.nih.gov/ij/ |
| LAS X | Leica | https://www.leica-microsystems.com/ |
| SlideBook 6.0 | Intelligent Imaging Innovations, Inc. | https://www.intelligent-imaging.com/slidebook |
| MAXCHELATOR | Chris Patton: cpatton@stanford.edu | https://somapp.ucdmc.ucdavis.edu/pharmacology/bers/maxchelator/ |
| Image Studio Lite | LI-COR | https://www.licor.com/bio/image-studio-lite/download |