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
|