KEY RESOURCES TABLE
REAGENT or RESOURCE | SOURCE | IDENTIFIER |
---|---|---|
Antibodies | ||
rabbit polyclonal anti-HIF1α | Novus Biologicals | Cat#NB100-479 |
rabbit polyclonal anti–calpain-2 large subunit | Cell Signaling | Cat#2539 |
chicken polyclonal anti-β-actin | Abcam | Cat#13822 |
rabbit polyclonal anti-β-actin | Cell Signaling | Cat#4967 |
rabbit polyclonal anti–talin-1 | Sigma Aldrich | Cat#T3287, Clone8d4 |
rabbit monoclonal anti-mouse CD29 | Millipore / Abcam | Cat#04-1109, clone EP1041Y / ab179471, clone 1A27 |
mouse monoclonal anti-CD29 | Merck Millipore | Cat#MAB2079Z, clone HUTS-4 |
mouse monoclonal anti-CD29 | Beckman Coulter | Cat#6603113, clone 4B4 |
rat monoclonal anti-CD29 | BD Biosciences | Cat#553715, clone 9EG7 |
rat IgG2a | BD Biosciences | Cat#553927, clone R5-95 |
armenian hamster monoclonal anti-CD29 (FITC-conjugated) | BioLegend | Cat#102206, clone HMβ1-1 |
armenian hamster IgG (FITC-conjugated) | BioLegend | Cat#400906, clone HTK888 |
rabbit monoclonal anti-cleaved caspase-3 | Cell Signaling | Cat#9664 |
mouse anti-CD29 | BioLegend | Cat#303010, cloneTS2/16 |
goat anti-rabbit (conjugated to horseradish peroxidase) | Jackson | Cat#211-032-171 |
goat anti-mouse (conjugated to horseradish peroxidase) | Jackson | Cat#115-0450174 |
goat anti-chicken (conjugated to horseradish peroxidase) | Jackson | Cat#103-035-155 |
rabbit-on-rodent HRP-polymer | Biocare Medical | Cat#RMR622 |
goat anti-rat IgG AF488 | Thermo Fisher Scientific | Cat#A21247 |
goat anti-rabbit IgG AF633 | Thermo Fisher Scientific | Cat#A21071 |
IRDye® 680RD Goat anti-Mouse IgG Secondary Antibody | Licor | 926-68070 |
IRDye® 680RD Goat anti-Rabbit IgG Secondary Antibody | Licor | 926-68071 |
Goat anti-Rat IgG Secondary Antibody Alexa Fluor 680 | Thermo Fisher Scientific | Cat#A21096 |
Biological samples | ||
Human HN-31 tumor xenograft | Jeffrey Myers Laboratory, MD Anderson Cancer Center | N/A |
Human Detroit-261 tumor xenograft | Jeffrey Myers Laboratory, MD Anderson Cancer Center | N/A |
Chemicals, peptides, and recombinant proteins | ||
Dimethyl oxalylglycine (DMOG) | Cayman Chemical | 71210-100 |
PD150606 | Cayman Chemical / Sigma Aldrich | 13859, CAS: 179528-45-1 / D5946 |
Etoposide | Sigma Aldrich | E1383, CAS: 33419-42-0 |
Oligomycin A | Selleckchem | S1478 |
2-Deoxy-D-glucose | Sigma Aldrich | D8375, CAS: 154-17-6 |
Carbonyl cyanide-4-(trifluoromethoxy)phenylhydrazone (FCCP) | Sigma Aldrich | C2920-10MG |
Rotenone | Sigma Aldrich | R8875, CAS: 83-79-4 |
Antimycin-a | Sigma Aldrich | A8674, CAS: 1397-94-0 |
Tetraethylbenzimidazolylcarbocyanine iodide (JC-1) | Sigma Aldrich / Enzo LifeSciences | T4069-5MG / ENZ-52304 |
t-BOC-L-leucyl-L-methionine amide (CMAC) | Thermo Fischer Scientific | A6520 |
Methyl cellulose | Sigma Aldrich | M6385 |
Collagen I, High Concentration, Rat Tail, 100mg |
Corning | Cat#354249 |
Bovine Type I Atelo-Collagen Solution, 3 mg/ml |
Advanced Biomatrix | Cat#5005-100ML |
LIVE/DEAD fixable stain | Thermo Fisher Scientific | Cat#L34976 |
MISSION siRNA transfection reagent | Sigma Aldrich | S1452 |
OptiMEM | Thermo Fisher Scientific | Cat# 31985062 |
Lipofectamine 2000 | Thermo Fisher Scientific | Cat# 11668019 |
Rodent Decloaker | Biocare Medical | SKU: RD913 |
Sodium oxamate | Sigma Aldrich | CAS: 565-73-1 |
Protease inhibitor cocktail | Roche | 926-68071 |
Novex™ 4–20% Tris-Glycine Mini Gels | Thermo Fisher Scientific | XP04200BOX |
cOmplete ULTRA Tablets (Protease inhibitor cocktail) |
Roche | 05892791001 |
PhosSTOP | Sigma Aldrich | PHOSS-RO |
Immobilon-PSQ Membrane, PVDF | Merck Millipore | Cat#ISEQ85R |
Bradford Reagent | Sigma Aldrich | B6916 |
Phalloidin | Thermo Fisher Scientific | A22283 |
DAPI | Thermo Fisher Scientific | D21490 |
Critical commercial assays | ||
Lactate Colorimetric/Fluorometric Assay Kit | BioVision | K607 |
Experimental models: Cell lines | ||
Mouse breast carcinoma: 4T1 cells | ATCC | CRL-2539; STR with matching profile; no interspecies cross contamination |
Human head and neck squamous carcinoma: UT-SCC38 cells | Turku University and Turku University Hospital, 20521 Turku, Finland | STR with unique profile; no interspecies cross contamination |
Human head and neck squamous carcinoma: UT-SCC58 cells | Obtained from Raider Grenman, University of Turku, Finland | STR with unique profile; no interspecies cross contamination |
Human head and neck squamous carcinoma: UT-SCC38 Lifeact-GFP cells | Friedl Laboratory | N/A |
Human head and neck squamous carcinoma: UT-SCC58 Lifeact-GFP cells | Friedl Laboratory | N/A |
Human head and neck squamous carcinoma: Detroit 562 cells | ATCC | CCL-138 |
Human head and neck squamous carcinoma: HN-31 cells | Provided by Dr. John Ensley, Wane State University, Detroit, MI | N/A |
Experimental models: Organisms/strains | ||
Swiss nu/nu nudes | MD Anderson Housing and Breeding Center in the Experimental Radiation Oncology Department, MD Anderson Cancer Center | N/A |
Athymic nude | ENVIGO | 069 |
Oligonucleotides | ||
MISSION siRNA targeting mouse CAPN2 (siRNA ID-#1): SASI_Mm01_00183464, sequence start 1243 | Sigma Aldrich | Cat#NM_009794 |
MISSION siRNA targeting mouse CAPN2 (siRNA ID-#2): SASI_Mm01_00183465, sequence start 1820, | Sigma Aldrich | Cat#NM_009794 |
MISSION siRNA targeting mouse CAPN2 (siRNA ID-#3): SASI_Mm01_00183467, sequence start 2132 | Sigma Aldrich | Cat#NM_009794 |
MISSION siRNA targeting human CAPN2 (siRNA ID-#1): SASI_Hs01_00059357, Sequence start 1053, | Sigma Aldrich | Cat#NM_001748 |
MISSION siRNA targeting human CAPN2 (siRNA ID-#2): SASI_Hs01_00059364, Sequence start 810 | Sigma Aldrich | Cat#NM_001748 |
MISSION siRNA targeting human CAPN2 (siRNA ID-#3): SASI_Hs01_00059356, Sequence start 2234 | Sigma Aldrich | Cat#NM_001748 |
MISSION siRNA universal negative control | Sigma Aldrich | Cat#SIC001 |
Recombinant DNA | ||
pEGFP N1 | ClonTech | Cat#6085-1 |
plasmid DNA construct GFP-TalinL432G | Addgene | Cat#26725 |
Software and algorithms | ||
Fiji | ImageJ | https://imagej.net/software/fiji/ |
GraphPad Prism 8 | GraphPad | https://www.graphpad.com/scientific-software/prism/ |
Autozell version 080912 | Center for Computing and Communication Technologies [TZI], University of Bremen, Germany | Proprietary code; can be obtained on specific request |
MATLAB | MathWorks | https://www.mathworks.com/products/matlab.html. |
Python version 3.8. | Python Software Foundation | https://www.python.org/ |
Matplotlib Version 3.4.3. | Matplotlib package | https://matplotlib.org/. |
Phase Field 2D Cell Motility Simulation v1.0. | Center for Theoretical Physical Biology, Rice University / Northeastern University, USA | https://github.com/yanjun07/phaseField |