Skip to main content
[Preprint]. 2023 Oct 3:2023.10.02.560521. [Version 1] doi: 10.1101/2023.10.02.560521

Key resource table:

Reagent or Resources Source Identifier
Antibodies
ATF6 Abcam Cat# ab122897, RRID:AB_10899171
ATF4 Cell Signaling Technology Cat# 11815, RRID:AB_2616025
GLUT1 Cell Signaling Technology Cat# 12939, RRID:AB_2687899
elF2α Cell Signaling Technology Cat# 9722, RRID:AB_2230924
p-elF2α Cell Signaling Technology Cat# 3398, RRID:AB_2096481)
IRE1α Cell Signaling Technology Cat# 3294, RRID:AB_823545
BIP Cell Signaling Technology Cat# 3177, RRID:AB_2119845
PERK Cell Signaling Technology Cat# 3192, RRID:AB_2095847
pPERK (Thr980) Cell Signaling Technology Cat# 3179, RRID:AB_2095853
cMyc Cell Signaling Technology Cat# 5605, RRID:AB_1903938
cMyc Bio-Rad Cat# VPA00575,
p-cMyc Cell Signaling Technology Cat# 13748, RRID:AB_2687518
MFN2 Cell Signaling Technology Cat# 11925, RRID:AB_2750893
PPARɣ Cell Signaling Technology Cat# 2435, RRID:AB_2166051
PGC1α Cell Signaling Technology Cat# 2178, RRID:AB_823600
ERK1/2 Cell Signaling Technology Cat# 9102, RRID:AB_330744
pERK1/2 Cell Signaling Technology Cat# 9101, RRID:AB_331646
OXPHOS Abcam Cat# ab110413, RRID:AB_2629281
STIM1 Cell Signaling Technology Cat# 4916; RRID:AB_2271287
STIM2 Cell Signaling Technology Cat# 4917; RRID:AB_2198021
IP3R1 Yule Lab NA
IP3R2 Yule Lab NA
IP3R3 BD Biosciences Cat# 610313; RRID:AB_397705
SERCA2 Cell Signaling Technology Cat# 4388S; RRID:AB_2227684
GAPDH Millipore Sigma Cat# MAB374, RRID:AB_2107445
IRDye 800CW Goat anti-Mouse Li-Core Biosciences Cat# 925–32210, RRID:AB_2687825
IRDye 800CW Donkey anti-Rabbit Li-Core Biosciences Cat# 925–32213, RRID:AB_2715510
IRDye 680RD Goat anti-Mouse Li-Core Biosciences Cat# 925–68070, RRID:AB_2651128
IRDye 680RD Goat anti-Rabbit Li-Core Biosciences Cat# 925–68071, RRID:AB_2721181
Chemicals
Mito Tracker Green FM Thermo Fisher Scientific Cat# M7514
Mito SOX Thermo Fisher Scientific Cat# M36008
TMRE Thermo Fisher Scientific T669
CellROX Thermo Fisher Scientific C10492
Fura-2 AM Thermo Fisher Scientific Cat# F1221
FBS VWR Cat# MISC-LFSCI-STP
Antibiotic and Antimycotic Thermo Fisher Scientific Cat# 15240062
McCoy’s 5A Corning Cat# 10–050CV
RPMI-1640 Corning Cat# 10–040CV
Lipofectamine 2000 Thermo Fisher Scientific Cat# 11668019
TrypLE Thermo Fisher Scientific Cat# 12605028
Proteinase K Viagen Biotech Cat# 501-PK
Direct PCR lysis reagent Viagen Biotech Cat# 302-C
Glucose Sigma-Aldrich Cat# D9434
Puromycin MP Biomedical Cat# 02100552
RIPA buffer Sigma Cat# R0278
ATP Sigma Cat# A9187
Cyclopiazonic Acid Alomone Labs Cat# C-750
Dextrose Fisher Scientific Cat# D14
Tris Base Fisher Scientific Cat# BP152–5
NaCl Fisher Scientific Cat# S671
MOPS SDS running buffer Thermo Fisher Scientific Cat# NP0001
Tris Glycine transfer buffer Bio-rad Cat#161–0734
KCl Fisher Scientific Cat# P217
MgCl2 Fisher Scientific Cat# M33
CaCl2 Fisher Scientific Cat# C614
HEPES Fisher Scientific Cat# BP310
LDS sample buffer Thermo Fisher Scientific Cat# NP0007
NuPAGE Bis-Tris precast gels Thermo Fisher Scientific Cat# NP0321
Polyvinylidene difluoride membrane Li-Core Biosciences Cat# 88518
Odyssey Blocking Buffer (TBS) Li-Core Biosciences Cat# 937–50003
Dextran sulfate sodium MP Biomedical Cat# 0216011080
FluoroBlok Corning Cat# 351152
BioCoat Tumor Invasion Plate Corning Cat# 80774380
BCA assay kit Thermo Fisher Scientific Cat# A53225
CyQUANT Thermo Fisher Scientific Cat# C35006
cDNA Reverse Transcription Kit Applied biosystems Cat# 4368814
DNase I Thermo Fisher Scientific Cat# 18068–015
TRIzol Thermo Fisher Scientific Cat# 15596018
SYBER select master mix Thermo Fisher Scientific Cat# 4472920
Seahorse XF DMEM Medium pH 7.4 Agilent Technologies Cat# 103575–100
Seahorse XF 100 mM pyruvate solution Agilent Technologies Cat# 103578–100
Seahorse XF 200 mM glutamine solution Agilent Technologies Cat# 103579–100
Seahorse XF 1.0 M glucose solution Agilent Technologies Cat# 103577–100
Seahorse XFp Mito Fuel Flex Test Kit Agilent Technologies Cat# 103270–100
Seahorse XFp Glycolysis Stress Test Kit Agilent Technologies Cat# 103017–100
Seahorse XFp Cell Mito Stress Test Kit Agilent Technologies Cat# 103010–100
Tris Glycine SDS Running Buffer (10X) Thermo Fisher Scientific Cat# LC2675
Novex 10% Zymogram Plus (Gelatin) Protein Gels, 1.0 mm, 10-well Thermo Fisher Scientific Cat# ZY00100
SimplyBlue Safe Stain Thermo Fisher Scientific Cat# LC6060
Tween 20 Fisher Scientific Cat# BP337
Cell Lines
Human colon cancer HCT116 cells (male) ATCC Cat# CCL-247, RRID:CVCL_0291
Human colon cancer DLD1 cells (male) ATCC Cat# CCL-221, RRID:CVCL_0248
Human colon cancer HCT116 STIM1 KO cells This paper N/A
Human colon cancer HCT116 STIM2 KO cells This paper N/A
Human colon cancer HCT116 DKO cells This paper N/A
Human colon cancer DLD1 STIM1 KO cells This paper N/A
Human colon cancer DLD1 STIM2 KO cells This paper N/A
Human colon cancer DLD1 DKO cells This paper N/A
Human colon cancer HCT116 Orai1 KO cells This paper N/A
Human colon cancer HCT116 TKO cells This paper N/A
Primer Sequences for RT-qPCR
MMP14- Forward 5’-AAGAGGAGAAGAGCAAACAG-3’
MMP14- Reverse 5’-CGGTAGGCACTGAACTTG-3’
MMP16- Forward 5’-AGCACTGGAAGACGGTTGG-3’
MMP16- Reverse 5’-CTCCGTTCCGCAGACTGTA-3’
SERCA1- Forward 5’-GTGATCCGCCAGCTAATG-3’
SERCA1- Reverse 5’-CGAATGTCAGGTCCGTCT-3’
SERCA2- Forward 5’-CGCTACCTCATCTCGTCCA-3’
SERCA2- Reverse 5’-TCGGGTATGGGGATTCAA-3’
SERCA3- Forward 5’-GATGGAGTGAACGACGCA-3’
SERCA3- Reverse 5’-CCAGGTATCGGAAGAAGAG-3’
Orai1- Forward 5’-GATGAGCCTCAACGAGCACT-3’
Orai1- Reverse 5’-ATTGCCACCATGGCGAAGC-3’
Orai2- Forward 5’-TGGCGGAAGCTCTACCTGAG-3’
Orai2- Reverse 5’-CGGGTACTGGTACTGCGTC-3’
Orai3- Forward 5’- GAGTGACCACGAGTACCCACC −3’
Orai3- Reverse 5’- GGGTACCATGATGGCTGTGG −3’
GAPDH- Forward 5’-CCCTTCATTGACCTCAACTACA-3’
GAPDH- Reverse 5’-ATGACAAGCTTCCCGTTCTC-3’
Tubulin- Forward 5’-AGTCCAAGCTGGAGTTCTCTAT-3’
Tubulin- Reverse 5’-CAATCAGAGTGCTCCAGGGT-3’
mtDNAR1- Forward 5’-CGGGCTACTACAACCCTTCG-3’
mtDNAR1- Reverse 5’-GATGGTAGATGTGGCGGGTT-3’
mtDNAR2- Forward 5’-CTATCCTGCCCGCCATCATC-3’
mtDNAR2- Reverse 5’-AAGGGAGGGATCGTTGACCT-3’
mtDNAR3- Forward 5’-TCATCCAAACCCCCTGAAGC-3’
mtDNAR3- Reverse 5’-CGACTGTGAGTGCGTTCGTA-3’
genomicDNAR1- Forward 5’-GCAACTTAGAGGTGGGGAGC-3’
genomic DNAR1- Reverse 5’-AACAAGCTTTGAGTGCAAGAGA-3’
genomicDNAR2- Forward 5’-GGGTGGGGGAGAGATTACCT-3’
genomic DNAR2- Reverse 5’-CAATCTGCCCAGAGAAGCCA-3’
Primer sequences for guide RNA
ORAI1 g1 CRISPR/Cas9 Guide RNA 5′-GTTGCTCACCGCCTCGATGT-3′ Trebak Lab (Emrich et al., 2021) N/A
ORAI2 g1 CRISPR/Cas9 Guide RNA 5′-ACGACAGGGCCTGTACCGAG-3′ Trebak Lab (Emrich et al., 2021) N/A
ORAI2 g3 CRISPR/Cas9 Guide RNA 5′-CTCATGCGGGGACTCGCTGA-3′ Trebak Lab (Emrich et al., 2021) N/A
ORAI3 g2 CRISPR/Cas9 Guide RNA 5′-GTTCGTGCACCGCGGCTACC-3′ Trebak Lab (Emrich et al., 2021) N/A
ORAI3 g4 CRISPR/Cas9 Guide RNA 5′-CCAGAGACTGCACCGCTACG-3′ Trebak Lab (Emrich et al., 2021) N/A
STIM1 g3 CRISPR/Cas9 Guide RNA 5′-TGATGAGCTTATCCTCACCA-3′ Trebak Lab (Emrich et al., 2021) N/A
STIM2 g1 CRISPR/Cas9 Guide RNA 5′-AGATGGTGGAATTGAAGTAG-3′ Trebak Lab (Emrich et al., 2021) N/A
Recombinant DNA
LentiCRISPR v2 Addgene Cat#52961
pCMV R-CEPIA1er Addgene Cat#58216
pSpCas9(BB)-2A-GFP (PX458) Addgene Cat#48138
pU6-(Bbs1)-mCherry Addgene Cat#64324
Software
Origin 2019b Origin labs https://www.originlab.com/
Image J NIH https://imagej.nih.gov/ij/
Image Studio Lite LI-COR https://www.licor.com/bio/image-studio-lite/download
LAS X Leica https://www.leica-microsystems.com/
Bio render Bio render https://www.biorender.com