Virus:Article Title: Mitochondrial cristae-remodeling protein OPA1 in POMC neurons couples Ca 2+ homeostasis with adipose tissue lipolysis
Article Snippet: .. REAGENT or RESOURCE SOURCE IDENTIFIER Antibodies Alpha-MSH Millipore Cat# AB5087; RRID: AB_91683 Alpha-Tubulin Sigma-Aldrich Cat# T6199; RRID: AB_477583 Alexa Fluor 594 chicken anti-mouse ThermoFisher Cat# A-21201; RRID: AB_2535787 Alexa Fluor 488 chicken anti-rabbit ThermoFisher Cat# A-21441; RRID: AB_2535859 Alexa Fluor 488 donkey anti-sheep ThermoFisher Cat# A-11015; RRID: AB_2534082 Alexa Fluor 594 donkey anti-rabbit ThermoFisher Cat# A-21207; RRID: AB_141637 Alexa Fluor 647 donkey anti-rabbit ThermoFisher Cat# A32795; RRID: AB_2762835 Anti-mouse HRP GE Healthcare UK Limited Cat# NA931; RRID: AB_772210 Anti-Rabbit HRP GE Healthcare UK Limited Cat# NA934; RRID: AB_2722659 b-Actin Sigma-Aldrich Cat# A2066; RRID: AB_476693 GFP Aves Labs Cat# GFP-1010; RRID: AB_2307313 HA.11 Covance Cat# MMS-101R; RRID: AB_291262 HSL-PSer563 Cell Signaling Cat# 4139; RRID: AB_2135495 HSL-PSer660 Cell Signaling Cat# 4126; RRID: AB_490997 HSL Cell Signaling Cat# 4107; RRID: AB_2296900 OPA1 BDBiosciences Cat# 612606; RRID: AB_399888 OPA1 Novus Biologicals Cat# NB110-55290C; RRID: AB_1851209 OXPHOS Rodent Antibody Cocktail Abcam Cat# ab110413; RRID: AB_2629281 Perilipin Cell Signaling Cat# 3467; RRID: AB_2167270 POMC Phoenix Pharmaceuticals Cat# H-029-30; RRID: AB_2307442 TOM20 Santa Cruz Cat# sc-11415; RRID: AB_2207533 Bacterial and virus strains AAV8-hSYN-DIO-hM3D(Gq)-mCherry Addgene Cat# 44361 AAV8-hSYN-DIO-hM4D(Gi)-mCherry Addgene Cat# 44362 pGP-Syn-FLEX-jGCaMP7s-WPRE Addgene Cat# 10449-AAV9 pAAV-FLEX-EF1A-mtGCaMP6s-WPRE Vector builder This paper pAAV-FLEX-tdTomato Addgene Cat# 28306-AAV9 Chemicals, peptides, and recombinant proteins 3-3’-Diaminobenzidine Sigma-Aldrich Cat# D5637 Adenosine diphosphate (ADP) Calbiochem Cat# 117105 Alpha-MSH Sigma-Aldrich Cat# M4135 Angiotensin II Sigma-Aldrich Cat# A9525 Antimycin A Sigma-Aldrich Cat# A8674 Artificial cerebrospinal fluid (aCSF) Tocris Bioscience Cat# 3525 Buffer RLT Qiagen Cat# 79216 Carbonyl cyanide 4-(trifluoromethoxy) phenylhydrazone (FCCP) Sigma-Aldrich Cat# C2920 Clozapine-N-oxide Tocris Cat# C4936 Dental acrylic TAB2000 , liquid Kerr Cat#61775 Dental acrylic TAB2000 , powder Kerr Cat#61771 Digitonin Sigma-Aldrich Cat# D5628 Durcupan Electron Microscopy Sciences Cat# 14040 Dynabeads Protein G ThermoFisher Cat# 10004D Enhanced chemiluminescence reagent Pierce Cat# 32105 (Continued on next page) e1 Cell Metabolism 33, 1820–1835.e1–e9, September 7, 2021 .. REAGENT or RESOURCE SOURCE IDENTIFIER Glucose solution 40% Fresenius Kabi Cat# C.N 620724 Glutamate Sigma-Aldrich Cat# G1626 Glutaraldehyde solution 50 % Panreac AppliChem Cat# A3166 Insulin Lilly Cat# C.N 710008.9 Kaempferol Sigma-Aldrich Cat# 1354900 Leptin R&D Cat# 498-OB Malate Sigma-Aldrich Cat# M1000 Melanotan II Bachem Cat# 4039778 Mitoxantrone Hydrochloride Sigma-Aldrich Cat# M2305000 Osmium tetroxide Electron Microscopy Sciences Cat# 19190 Paraformaldehyde 16% Electron Microscopy Sciences Cat# 15710-S Picric acid solution Sigma-Aldrich Cat# P6744 Protease and Phosphatase Inhibitor Cocktail Sigma-Aldrich Cat# PPC1010 RIPA Buffer Sigma-Aldrich Cat# R0278 Rotenone Sigma-Aldrich Cat# R8875 Ru265 Dr. Justin J Wilson N/A Ru360 MerkMillipore Cat# 557440 SB202190 Tocris Cat# 1264 Sodium pyruvate Sigma-Aldrich Cat# P2256 SR59230A Tocris Cat# 1511 Succinate Sigma-Aldrich Cat# S2378 Sucrose Sigma-Aldrich Cat# S0389 Trizol Reagent ThermoFisher Cat# 15596026 Uranyl acetate Electron Microscopy Sciences Cat# 22400 Critical commercial assays Corticosterone EIA Immunodiagnosticsystems Cat# AC-14F1 ECL Western Blotting Substrate Pierce Cat# 32106 Epinephrine/Norepinephrine ELISA LDN Cat# BA E-5400 Glucocard X-meter Arkray Cat# GT-1910 High Capacity cDNA Reverse Transcription Kit Applied Biosystems Cat#4368814 HR Series NEFA-HR(2)-R1 WAKO Cat# 434-91795 HR Series NEFA-HR(2)-R2 WAKO Cat# 436-91995 Insulin ELISA CrystalChem Cat# 90080 Leptin ELISA CrystalChem Cat# 90030 MSH, alpha EIA Phoenix Pharmaceuticals Cat# EK-043-01 MSH, alpha FIA Phoenix Pharmaceuticals Cat# FEK-043-01 PreMix Ex taq Takara C#RR39WR Quant-iT RiboGreen RNA assay kit ThermoFisher Cat# R1149 RNeasy-plus Mini kit Qiagen Cat# 74134 RNAscope Probe Diluent ACD; Advanced Cell Diagnostics Cat# 3000041 RNAscope Fluorescent Multiplex Detection Reagents ACD; Advanced Cell Diagnostics Cat# 320851 RNAscope Enhancer Fluorescent Kit v2 ACD; Advanced Cell Diagnostics Cat# 323100 RNAscope HybEz hybridization system Advanced Cell Diagnostics Cat# 311720 SDS-PAGE pre-cast 4%-12% gels Bio-Rad Cat# 345-0124 Triglyceride determination kit Sigma-Aldrich Cat# TR0100-1KT Experimental models: Organisms/strains POMC-Cre mice (Xu et al., 2005) N/A B6N.129-Rpl22tm1.1Psam/J mice The Jackson Laboratory Cat# 011029 Opa1fl/fl mice (Ramı́rez et al., 2017) N/A (Continued on next page) ll OPEN ACCESSArticle Cell Metabolism 33, 1820–1835.e1–e9, September 7, 2021 e2
Recombinant:Article Title: Mitochondrial cristae-remodeling protein OPA1 in POMC neurons couples Ca 2+ homeostasis with adipose tissue lipolysis
Article Snippet: .. REAGENT or RESOURCE SOURCE IDENTIFIER Antibodies Alpha-MSH Millipore Cat# AB5087; RRID: AB_91683 Alpha-Tubulin Sigma-Aldrich Cat# T6199; RRID: AB_477583 Alexa Fluor 594 chicken anti-mouse ThermoFisher Cat# A-21201; RRID: AB_2535787 Alexa Fluor 488 chicken anti-rabbit ThermoFisher Cat# A-21441; RRID: AB_2535859 Alexa Fluor 488 donkey anti-sheep ThermoFisher Cat# A-11015; RRID: AB_2534082 Alexa Fluor 594 donkey anti-rabbit ThermoFisher Cat# A-21207; RRID: AB_141637 Alexa Fluor 647 donkey anti-rabbit ThermoFisher Cat# A32795; RRID: AB_2762835 Anti-mouse HRP GE Healthcare UK Limited Cat# NA931; RRID: AB_772210 Anti-Rabbit HRP GE Healthcare UK Limited Cat# NA934; RRID: AB_2722659 b-Actin Sigma-Aldrich Cat# A2066; RRID: AB_476693 GFP Aves Labs Cat# GFP-1010; RRID: AB_2307313 HA.11 Covance Cat# MMS-101R; RRID: AB_291262 HSL-PSer563 Cell Signaling Cat# 4139; RRID: AB_2135495 HSL-PSer660 Cell Signaling Cat# 4126; RRID: AB_490997 HSL Cell Signaling Cat# 4107; RRID: AB_2296900 OPA1 BDBiosciences Cat# 612606; RRID: AB_399888 OPA1 Novus Biologicals Cat# NB110-55290C; RRID: AB_1851209 OXPHOS Rodent Antibody Cocktail Abcam Cat# ab110413; RRID: AB_2629281 Perilipin Cell Signaling Cat# 3467; RRID: AB_2167270 POMC Phoenix Pharmaceuticals Cat# H-029-30; RRID: AB_2307442 TOM20 Santa Cruz Cat# sc-11415; RRID: AB_2207533 Bacterial and virus strains AAV8-hSYN-DIO-hM3D(Gq)-mCherry Addgene Cat# 44361 AAV8-hSYN-DIO-hM4D(Gi)-mCherry Addgene Cat# 44362 pGP-Syn-FLEX-jGCaMP7s-WPRE Addgene Cat# 10449-AAV9 pAAV-FLEX-EF1A-mtGCaMP6s-WPRE Vector builder This paper pAAV-FLEX-tdTomato Addgene Cat# 28306-AAV9 Chemicals, peptides, and recombinant proteins 3-3’-Diaminobenzidine Sigma-Aldrich Cat# D5637 Adenosine diphosphate (ADP) Calbiochem Cat# 117105 Alpha-MSH Sigma-Aldrich Cat# M4135 Angiotensin II Sigma-Aldrich Cat# A9525 Antimycin A Sigma-Aldrich Cat# A8674 Artificial cerebrospinal fluid (aCSF) Tocris Bioscience Cat# 3525 Buffer RLT Qiagen Cat# 79216 Carbonyl cyanide 4-(trifluoromethoxy) phenylhydrazone (FCCP) Sigma-Aldrich Cat# C2920 Clozapine-N-oxide Tocris Cat# C4936 Dental acrylic TAB2000 , liquid Kerr Cat#61775 Dental acrylic TAB2000 , powder Kerr Cat#61771 Digitonin Sigma-Aldrich Cat# D5628 Durcupan Electron Microscopy Sciences Cat# 14040 Dynabeads Protein G ThermoFisher Cat# 10004D Enhanced chemiluminescence reagent Pierce Cat# 32105 (Continued on next page) e1 Cell Metabolism 33, 1820–1835.e1–e9, September 7, 2021 .. REAGENT or RESOURCE SOURCE IDENTIFIER Glucose solution 40% Fresenius Kabi Cat# C.N 620724 Glutamate Sigma-Aldrich Cat# G1626 Glutaraldehyde solution 50 % Panreac AppliChem Cat# A3166 Insulin Lilly Cat# C.N 710008.9 Kaempferol Sigma-Aldrich Cat# 1354900 Leptin R&D Cat# 498-OB Malate Sigma-Aldrich Cat# M1000 Melanotan II Bachem Cat# 4039778 Mitoxantrone Hydrochloride Sigma-Aldrich Cat# M2305000 Osmium tetroxide Electron Microscopy Sciences Cat# 19190 Paraformaldehyde 16% Electron Microscopy Sciences Cat# 15710-S Picric acid solution Sigma-Aldrich Cat# P6744 Protease and Phosphatase Inhibitor Cocktail Sigma-Aldrich Cat# PPC1010 RIPA Buffer Sigma-Aldrich Cat# R0278 Rotenone Sigma-Aldrich Cat# R8875 Ru265 Dr. Justin J Wilson N/A Ru360 MerkMillipore Cat# 557440 SB202190 Tocris Cat# 1264 Sodium pyruvate Sigma-Aldrich Cat# P2256 SR59230A Tocris Cat# 1511 Succinate Sigma-Aldrich Cat# S2378 Sucrose Sigma-Aldrich Cat# S0389 Trizol Reagent ThermoFisher Cat# 15596026 Uranyl acetate Electron Microscopy Sciences Cat# 22400 Critical commercial assays Corticosterone EIA Immunodiagnosticsystems Cat# AC-14F1 ECL Western Blotting Substrate Pierce Cat# 32106 Epinephrine/Norepinephrine ELISA LDN Cat# BA E-5400 Glucocard X-meter Arkray Cat# GT-1910 High Capacity cDNA Reverse Transcription Kit Applied Biosystems Cat#4368814 HR Series NEFA-HR(2)-R1 WAKO Cat# 434-91795 HR Series NEFA-HR(2)-R2 WAKO Cat# 436-91995 Insulin ELISA CrystalChem Cat# 90080 Leptin ELISA CrystalChem Cat# 90030 MSH, alpha EIA Phoenix Pharmaceuticals Cat# EK-043-01 MSH, alpha FIA Phoenix Pharmaceuticals Cat# FEK-043-01 PreMix Ex taq Takara C#RR39WR Quant-iT RiboGreen RNA assay kit ThermoFisher Cat# R1149 RNeasy-plus Mini kit Qiagen Cat# 74134 RNAscope Probe Diluent ACD; Advanced Cell Diagnostics Cat# 3000041 RNAscope Fluorescent Multiplex Detection Reagents ACD; Advanced Cell Diagnostics Cat# 320851 RNAscope Enhancer Fluorescent Kit v2 ACD; Advanced Cell Diagnostics Cat# 323100 RNAscope HybEz hybridization system Advanced Cell Diagnostics Cat# 311720 SDS-PAGE pre-cast 4%-12% gels Bio-Rad Cat# 345-0124 Triglyceride determination kit Sigma-Aldrich Cat# TR0100-1KT Experimental models: Organisms/strains POMC-Cre mice (Xu et al., 2005) N/A B6N.129-Rpl22tm1.1Psam/J mice The Jackson Laboratory Cat# 011029 Opa1fl/fl mice (Ramı́rez et al., 2017) N/A (Continued on next page) ll OPEN ACCESSArticle Cell Metabolism 33, 1820–1835.e1–e9, September 7, 2021 e2
Electron Microscopy:Article Title: Mitochondrial cristae-remodeling protein OPA1 in POMC neurons couples Ca 2+ homeostasis with adipose tissue lipolysis
Article Snippet: .. REAGENT or RESOURCE SOURCE IDENTIFIER Antibodies Alpha-MSH Millipore Cat# AB5087; RRID: AB_91683 Alpha-Tubulin Sigma-Aldrich Cat# T6199; RRID: AB_477583 Alexa Fluor 594 chicken anti-mouse ThermoFisher Cat# A-21201; RRID: AB_2535787 Alexa Fluor 488 chicken anti-rabbit ThermoFisher Cat# A-21441; RRID: AB_2535859 Alexa Fluor 488 donkey anti-sheep ThermoFisher Cat# A-11015; RRID: AB_2534082 Alexa Fluor 594 donkey anti-rabbit ThermoFisher Cat# A-21207; RRID: AB_141637 Alexa Fluor 647 donkey anti-rabbit ThermoFisher Cat# A32795; RRID: AB_2762835 Anti-mouse HRP GE Healthcare UK Limited Cat# NA931; RRID: AB_772210 Anti-Rabbit HRP GE Healthcare UK Limited Cat# NA934; RRID: AB_2722659 b-Actin Sigma-Aldrich Cat# A2066; RRID: AB_476693 GFP Aves Labs Cat# GFP-1010; RRID: AB_2307313 HA.11 Covance Cat# MMS-101R; RRID: AB_291262 HSL-PSer563 Cell Signaling Cat# 4139; RRID: AB_2135495 HSL-PSer660 Cell Signaling Cat# 4126; RRID: AB_490997 HSL Cell Signaling Cat# 4107; RRID: AB_2296900 OPA1 BDBiosciences Cat# 612606; RRID: AB_399888 OPA1 Novus Biologicals Cat# NB110-55290C; RRID: AB_1851209 OXPHOS Rodent Antibody Cocktail Abcam Cat# ab110413; RRID: AB_2629281 Perilipin Cell Signaling Cat# 3467; RRID: AB_2167270 POMC Phoenix Pharmaceuticals Cat# H-029-30; RRID: AB_2307442 TOM20 Santa Cruz Cat# sc-11415; RRID: AB_2207533 Bacterial and virus strains AAV8-hSYN-DIO-hM3D(Gq)-mCherry Addgene Cat# 44361 AAV8-hSYN-DIO-hM4D(Gi)-mCherry Addgene Cat# 44362 pGP-Syn-FLEX-jGCaMP7s-WPRE Addgene Cat# 10449-AAV9 pAAV-FLEX-EF1A-mtGCaMP6s-WPRE Vector builder This paper pAAV-FLEX-tdTomato Addgene Cat# 28306-AAV9 Chemicals, peptides, and recombinant proteins 3-3’-Diaminobenzidine Sigma-Aldrich Cat# D5637 Adenosine diphosphate (ADP) Calbiochem Cat# 117105 Alpha-MSH Sigma-Aldrich Cat# M4135 Angiotensin II Sigma-Aldrich Cat# A9525 Antimycin A Sigma-Aldrich Cat# A8674 Artificial cerebrospinal fluid (aCSF) Tocris Bioscience Cat# 3525 Buffer RLT Qiagen Cat# 79216 Carbonyl cyanide 4-(trifluoromethoxy) phenylhydrazone (FCCP) Sigma-Aldrich Cat# C2920 Clozapine-N-oxide Tocris Cat# C4936 Dental acrylic TAB2000 , liquid Kerr Cat#61775 Dental acrylic TAB2000 , powder Kerr Cat#61771 Digitonin Sigma-Aldrich Cat# D5628 Durcupan Electron Microscopy Sciences Cat# 14040 Dynabeads Protein G ThermoFisher Cat# 10004D Enhanced chemiluminescence reagent Pierce Cat# 32105 (Continued on next page) e1 Cell Metabolism 33, 1820–1835.e1–e9, September 7, 2021 .. REAGENT or RESOURCE SOURCE IDENTIFIER Glucose solution 40% Fresenius Kabi Cat# C.N 620724 Glutamate Sigma-Aldrich Cat# G1626 Glutaraldehyde solution 50 % Panreac AppliChem Cat# A3166 Insulin Lilly Cat# C.N 710008.9 Kaempferol Sigma-Aldrich Cat# 1354900 Leptin R&D Cat# 498-OB Malate Sigma-Aldrich Cat# M1000 Melanotan II Bachem Cat# 4039778 Mitoxantrone Hydrochloride Sigma-Aldrich Cat# M2305000 Osmium tetroxide Electron Microscopy Sciences Cat# 19190 Paraformaldehyde 16% Electron Microscopy Sciences Cat# 15710-S Picric acid solution Sigma-Aldrich Cat# P6744 Protease and Phosphatase Inhibitor Cocktail Sigma-Aldrich Cat# PPC1010 RIPA Buffer Sigma-Aldrich Cat# R0278 Rotenone Sigma-Aldrich Cat# R8875 Ru265 Dr. Justin J Wilson N/A Ru360 MerkMillipore Cat# 557440 SB202190 Tocris Cat# 1264 Sodium pyruvate Sigma-Aldrich Cat# P2256 SR59230A Tocris Cat# 1511 Succinate Sigma-Aldrich Cat# S2378 Sucrose Sigma-Aldrich Cat# S0389 Trizol Reagent ThermoFisher Cat# 15596026 Uranyl acetate Electron Microscopy Sciences Cat# 22400 Critical commercial assays Corticosterone EIA Immunodiagnosticsystems Cat# AC-14F1 ECL Western Blotting Substrate Pierce Cat# 32106 Epinephrine/Norepinephrine ELISA LDN Cat# BA E-5400 Glucocard X-meter Arkray Cat# GT-1910 High Capacity cDNA Reverse Transcription Kit Applied Biosystems Cat#4368814 HR Series NEFA-HR(2)-R1 WAKO Cat# 434-91795 HR Series NEFA-HR(2)-R2 WAKO Cat# 436-91995 Insulin ELISA CrystalChem Cat# 90080 Leptin ELISA CrystalChem Cat# 90030 MSH, alpha EIA Phoenix Pharmaceuticals Cat# EK-043-01 MSH, alpha FIA Phoenix Pharmaceuticals Cat# FEK-043-01 PreMix Ex taq Takara C#RR39WR Quant-iT RiboGreen RNA assay kit ThermoFisher Cat# R1149 RNeasy-plus Mini kit Qiagen Cat# 74134 RNAscope Probe Diluent ACD; Advanced Cell Diagnostics Cat# 3000041 RNAscope Fluorescent Multiplex Detection Reagents ACD; Advanced Cell Diagnostics Cat# 320851 RNAscope Enhancer Fluorescent Kit v2 ACD; Advanced Cell Diagnostics Cat# 323100 RNAscope HybEz hybridization system Advanced Cell Diagnostics Cat# 311720 SDS-PAGE pre-cast 4%-12% gels Bio-Rad Cat# 345-0124 Triglyceride determination kit Sigma-Aldrich Cat# TR0100-1KT Experimental models: Organisms/strains POMC-Cre mice (Xu et al., 2005) N/A B6N.129-Rpl22tm1.1Psam/J mice The Jackson Laboratory Cat# 011029 Opa1fl/fl mice (Ramı́rez et al., 2017) N/A (Continued on next page) ll OPEN ACCESSArticle Cell Metabolism 33, 1820–1835.e1–e9, September 7, 2021 e2
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