Isolation:Article Title: Stromal lipid species dictate melanoma metastasis and tropism.
Article Snippet: .. REAGENT or RESOURCE SOURCE IDENTIFIER Antibodies Phospho-Akt (Ser473)(D9E)XP Rabbit mAb Cell Signaling Cat #4060; RRID: AB_2315049 Phospho-Akt (Thr308)(244F)Rabbit mAb Cell Signaling Cat #4056; RRID:AB_331163 Akt Antibody Cell Signaling Cat #9272; RRID:AB_329827 Phospho-mTOR (Ser2448)(D9C2)XP Rabbit mAb Cell Signaling Cat#5536; RRID:AB_10691552 mTOR Antibody Cell Signaling Cat#2972; RRID:AB_330978 Phospho-Stat3 (Tyr705) Antibody Cell Signaling Cat#9131; RRID:AB_331586 STAT3 Antibody Cell Signaling Cat#9132; RRID:AB_331588 BCL2 (124) Mouse mAb Cell Signaling Cat#15071; RRID:AB_2744528 Anti- beta Tubulin Antibody Abcam ab6046; RRID:AB_2210370 Anti- Cytochrome C Antibody [7H8.2C12] Abcam ab13575; RRID:AB_300470 CPT1A Antibody Proteintech Cat#66039-1; RRID:AB_11041710 Anti-TOMM20 Antibody [4F3]- BSA and Azide Free Abcam ab56783; RRID:AB_945896 Anti-Rabbit IgG, HRP-linked Antibody Cell Signaling Cat#7074; RRID:AB_2099233 Anti- Mouse IgG, HRP-linked Antibody Cell Signaling Cat#7076; RRID:AB_330924 Goat anti-Mouse Alexa Fluor 488 Thermo Fisher A32723; RRID:AB_2633275 Goat anti-Rabbit Alexa Fluor 555 Thermo Fisher A32732; RRID:AB_2633281 Fatty Acid Synthase (C20G5) Rabbit mAb Cell Signaling Cat #3180S; RRID:N/A CD36 (E8B7S)Rabbit mAb Cell Signaling Cat #28109S; RRID:N/A PPARg (C26H12) Rabbit mAb Cell Signaling Cat #2435s; RRID:N/A Anti- FABP4 Antibody [EPR3579] Abcam ab92501; RRID:AB_10562486 InVivoMAb anti-mouse IL-6R Bio X cell BE0047; RRID:AB_1107588 Biological samples Human Dog ears for Preadipocytes’ isolation This study Biobank Chemicals, peptides, and recombinant proteins Trypsin -EDTA (0.25%), phenol red Gibco 25200056 DMEM with glutamax Thermo Fisher 10566016 FBS Thermo Fisher A5256801 Penicillin – Streptomycin Thermo Fisher 15140122 Glutamax Thermo Fisher 35050061 Non essential Amino acids Thermo Fisher 11140050 Cleanscite Lipid Removal Kit Biotech Support Group X2555-100 Dispase II Gibco Cat no: 17105041 Collagenase I Gibco Cat no: 17018029 Adipocytes Maintenance media Zenbio AM-1 PGM-2 Preadipocyte Growth Medium -2 Bullekit Lonza PT 8002 BODIPY 493/503 Cayman CAS 121207-31-6 DMSO Sigma Aldrich D540 PBS Thermo Scientific J61196.AP Cy5 -PC Avanti Lipids 850483 Hoechst Thermo Fisher 62249 CellTracker Orange CMRA Invitrogen C34551 Mitoview 633 Biotium BT70055 MitoTracker Green Dyes Invitrogen M7514 Seahorse XF base medium Agilent 103335–100 (Continued on next page) e1 Cancer Cell 43, 1–17.e1–e11, June 9, 2025 .. REAGENT or RESOURCE SOURCE IDENTIFIER Glucose Solution Gibco A2494001 Sodium Pyruvate (100mM) Gibco 11360070 Triton X 100 Sigma Aldrich X100-500ML Sodium chloride Sigma Aldrich S6546 Tris Sigma Aldrich 93362 MOPS Sigma Aldrich M1254 Chloroform Thermo Fisher J67241.K2 Tween 20 Sigma Aldrich P1379 10X Tris Buffered Saline (TBS) Bio-Rad 1706435 Methanol Merck 1.06007 Acetone Sigma Aldrich 320110 UltraPure SDS Solution,10% Thermo Fisher 15553027 4% Paraformaldehyde in PBS Thermo Scientific J61899.AK 10%Formalin Sigma Aldrich HT501640-19L Tris- Glycine- SDS Buffer 10x Concentrate Sigma Aldrich T7777-1L Sucrose Sigma Aldrich S9378 Ethylene -bis (oxyethylenenitrilo)tetraacetic acid (EGTA) Sigma Aldrich ED2SS Beta- mercaptoethanol Sigma Aldrich M6250-M6250 Laemmli Buffer Bio-Rad 610747 Phosphatase Inhibitor Roche 4906845001 Protease Inhibitor Roche 11836153001 Mini-PROTEAN TGX gels Bio-Rad 4568084 Precision Plus Ladder Bio-Rad 1610374 1X TGS buffer Bio-Rad 1610732 Ponceau Stain G-Biosciences 786-575 TransBlot Tubro system Bio-Rad 170-4270 BSA Sigma A9418 ECL western blot substrate Promega w1001 Polybrene Santa Cruz Biotechnology sc-134220 Puromycin Sigma Aldrich P8833-10MG Egg-PC Avanti Polar Lipids 840051 Ceramide ThermoFisher D-7540 JR-AB2-011 MedChem Express HY-122022 LY294002 Cambridge Bioscience 70920-5mg-CAY ABT199 ApeXBio A8194 Salvianolic acid Cayman chemical 9001577 Etomoxir MedChemExpress HY-50202 Rotenone Sigma Aldrich R8875 Trizol solution Invitrogen 12044977 Antimycin A Sigma Aldrich A8674 Carbonyl cyanide 4-(trifluoromethoxy)phenylhydrazone Sigma Aldrich C920-10MG Oligomycin Sigma Aldrich 75351-5MG SYBR Green Sigma Aldrich 4913850001 CellROX Green Thermo Fisher C10444 MitoSOX Red mitochondrial superoxide indicator Thermo Fisher M36008 Collagenase IV Worthington Biochemical LS004186 ONC201 Apex Bio A8724 N-acetylcysteine Sigma Aldrich A0737 (Continued on next page) Cancer Cell 43, 1–17.e1–e11, June 9, 2025 e2
Recombinant:Article Title: Stromal lipid species dictate melanoma metastasis and tropism.
Article Snippet: .. REAGENT or RESOURCE SOURCE IDENTIFIER Antibodies Phospho-Akt (Ser473)(D9E)XP Rabbit mAb Cell Signaling Cat #4060; RRID: AB_2315049 Phospho-Akt (Thr308)(244F)Rabbit mAb Cell Signaling Cat #4056; RRID:AB_331163 Akt Antibody Cell Signaling Cat #9272; RRID:AB_329827 Phospho-mTOR (Ser2448)(D9C2)XP Rabbit mAb Cell Signaling Cat#5536; RRID:AB_10691552 mTOR Antibody Cell Signaling Cat#2972; RRID:AB_330978 Phospho-Stat3 (Tyr705) Antibody Cell Signaling Cat#9131; RRID:AB_331586 STAT3 Antibody Cell Signaling Cat#9132; RRID:AB_331588 BCL2 (124) Mouse mAb Cell Signaling Cat#15071; RRID:AB_2744528 Anti- beta Tubulin Antibody Abcam ab6046; RRID:AB_2210370 Anti- Cytochrome C Antibody [7H8.2C12] Abcam ab13575; RRID:AB_300470 CPT1A Antibody Proteintech Cat#66039-1; RRID:AB_11041710 Anti-TOMM20 Antibody [4F3]- BSA and Azide Free Abcam ab56783; RRID:AB_945896 Anti-Rabbit IgG, HRP-linked Antibody Cell Signaling Cat#7074; RRID:AB_2099233 Anti- Mouse IgG, HRP-linked Antibody Cell Signaling Cat#7076; RRID:AB_330924 Goat anti-Mouse Alexa Fluor 488 Thermo Fisher A32723; RRID:AB_2633275 Goat anti-Rabbit Alexa Fluor 555 Thermo Fisher A32732; RRID:AB_2633281 Fatty Acid Synthase (C20G5) Rabbit mAb Cell Signaling Cat #3180S; RRID:N/A CD36 (E8B7S)Rabbit mAb Cell Signaling Cat #28109S; RRID:N/A PPARg (C26H12) Rabbit mAb Cell Signaling Cat #2435s; RRID:N/A Anti- FABP4 Antibody [EPR3579] Abcam ab92501; RRID:AB_10562486 InVivoMAb anti-mouse IL-6R Bio X cell BE0047; RRID:AB_1107588 Biological samples Human Dog ears for Preadipocytes’ isolation This study Biobank Chemicals, peptides, and recombinant proteins Trypsin -EDTA (0.25%), phenol red Gibco 25200056 DMEM with glutamax Thermo Fisher 10566016 FBS Thermo Fisher A5256801 Penicillin – Streptomycin Thermo Fisher 15140122 Glutamax Thermo Fisher 35050061 Non essential Amino acids Thermo Fisher 11140050 Cleanscite Lipid Removal Kit Biotech Support Group X2555-100 Dispase II Gibco Cat no: 17105041 Collagenase I Gibco Cat no: 17018029 Adipocytes Maintenance media Zenbio AM-1 PGM-2 Preadipocyte Growth Medium -2 Bullekit Lonza PT 8002 BODIPY 493/503 Cayman CAS 121207-31-6 DMSO Sigma Aldrich D540 PBS Thermo Scientific J61196.AP Cy5 -PC Avanti Lipids 850483 Hoechst Thermo Fisher 62249 CellTracker Orange CMRA Invitrogen C34551 Mitoview 633 Biotium BT70055 MitoTracker Green Dyes Invitrogen M7514 Seahorse XF base medium Agilent 103335–100 (Continued on next page) e1 Cancer Cell 43, 1–17.e1–e11, June 9, 2025 .. REAGENT or RESOURCE SOURCE IDENTIFIER Glucose Solution Gibco A2494001 Sodium Pyruvate (100mM) Gibco 11360070 Triton X 100 Sigma Aldrich X100-500ML Sodium chloride Sigma Aldrich S6546 Tris Sigma Aldrich 93362 MOPS Sigma Aldrich M1254 Chloroform Thermo Fisher J67241.K2 Tween 20 Sigma Aldrich P1379 10X Tris Buffered Saline (TBS) Bio-Rad 1706435 Methanol Merck 1.06007 Acetone Sigma Aldrich 320110 UltraPure SDS Solution,10% Thermo Fisher 15553027 4% Paraformaldehyde in PBS Thermo Scientific J61899.AK 10%Formalin Sigma Aldrich HT501640-19L Tris- Glycine- SDS Buffer 10x Concentrate Sigma Aldrich T7777-1L Sucrose Sigma Aldrich S9378 Ethylene -bis (oxyethylenenitrilo)tetraacetic acid (EGTA) Sigma Aldrich ED2SS Beta- mercaptoethanol Sigma Aldrich M6250-M6250 Laemmli Buffer Bio-Rad 610747 Phosphatase Inhibitor Roche 4906845001 Protease Inhibitor Roche 11836153001 Mini-PROTEAN TGX gels Bio-Rad 4568084 Precision Plus Ladder Bio-Rad 1610374 1X TGS buffer Bio-Rad 1610732 Ponceau Stain G-Biosciences 786-575 TransBlot Tubro system Bio-Rad 170-4270 BSA Sigma A9418 ECL western blot substrate Promega w1001 Polybrene Santa Cruz Biotechnology sc-134220 Puromycin Sigma Aldrich P8833-10MG Egg-PC Avanti Polar Lipids 840051 Ceramide ThermoFisher D-7540 JR-AB2-011 MedChem Express HY-122022 LY294002 Cambridge Bioscience 70920-5mg-CAY ABT199 ApeXBio A8194 Salvianolic acid Cayman chemical 9001577 Etomoxir MedChemExpress HY-50202 Rotenone Sigma Aldrich R8875 Trizol solution Invitrogen 12044977 Antimycin A Sigma Aldrich A8674 Carbonyl cyanide 4-(trifluoromethoxy)phenylhydrazone Sigma Aldrich C920-10MG Oligomycin Sigma Aldrich 75351-5MG SYBR Green Sigma Aldrich 4913850001 CellROX Green Thermo Fisher C10444 MitoSOX Red mitochondrial superoxide indicator Thermo Fisher M36008 Collagenase IV Worthington Biochemical LS004186 ONC201 Apex Bio A8724 N-acetylcysteine Sigma Aldrich A0737 (Continued on next page) Cancer Cell 43, 1–17.e1–e11, June 9, 2025 e2
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