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<ol>
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</ol>

<table><tbody><tr><td>100 mm plate</td><td>SPL Life Science</td><td>10101SPL</td><td></td></tr><tr><td>10x Tris/Glycine Buffer</td><td>BioRad</td><td>1610734</td><td></td></tr><tr><td>12 mm glass coverslips</td><td>Marienfeld</td><td>111520</td><td></td></tr><tr><td>12-well plate</td><td>SPL Life Science</td><td>30012SPL</td><td></td></tr><tr><td>16% Paraformaldehyde Aqueous Solution</td><td>Invitrogen</td><td>28908</td><td></td></tr><tr><td>18 mm glass coverslips</td><td>HART</td><td>-</td><td></td></tr><tr><td>24-well plate</td><td>SPL Life Science</td><td>30024SPL</td><td></td></tr><tr><td>3,3&prime;,5-Triiodo-L-thyronine sodium salt (T3)</td><td>Sigma</td><td>T6397</td><td></td></tr><tr><td>3-Isobutyl-1-methylxanthine (IBMX)</td><td>Calbiochem</td><td>410957</td><td></td></tr><tr><td>30% Acrylamide/Bis Solution, 37.5:1</td><td>BioRad</td><td>1610158</td><td></td></tr><tr><td>35 mm plate</td><td>SPL Life Science</td><td>20035SPL</td><td></td></tr><tr><td>6-well plate</td><td>SPL Life Science</td><td>30006SPL</td><td></td></tr><tr><td>60 mm culture dish</td><td>SPL Life Science</td><td>20060SPL</td><td>Usado para experimentos primários de pré-adipócito e DNA/qPCR; fornecedor/catálogo não fornecido</td></tr><tr><td>AccuRuler RGB Plus Pre-stained Protein Ladder</td><td>Maestrogen</td><td>02102-250</td><td></td></tr><tr><td>ACK lysing buffer</td><td>Gibco</td><td>A10492-01</td><td></td></tr><tr><td>Actrapid 100 UI/mL (Insulina)</td><td>Novo Nordisk</td><td>A10AB01</td><td></td></tr><tr><td>ADP</td><td>Merck</td><td>117105</td><td></td></tr><tr><td>Agarose</td><td>FERMELO BIOTEC</td><td>FER00AL200G</td><td>Usado para verificação de integridade do RNA; fornecedor/catálogo não fornecido</td></tr><tr><td>Ammonium Persulfate</td><td>BioRad</td><td>1610700</td><td></td></tr><tr><td>Anti-rabbit IgG, HRP-linked Antibody</td><td>Cell Signaling</td><td>7074</td><td></td></tr><tr><td>Anti-rat IgG, HRP-linked Antibody</td><td>Cell Signaling</td><td>7077</td><td></td></tr><tr><td>Antibiotic-antimycotic</td><td>Gibco</td><td>15240062</td><td></td></tr><tr><td>Axio Observer inverted microscope stand</td><td>Carl Zeiss</td><td>431007-9901-000</td><td>Adicionado dos Métodos Suplementares</td></tr><tr><td>BCA Protein Assay Kit</td><td>Thermo Scientific</td><td>D49328</td><td></td></tr><tr><td>BD Cytometer Setup and Tracking beads</td><td>BD Biosciences</td><td>-</td><td>Adicionado dos Métodos Suplementares; catálogo não fornecido</td></tr><tr><td>BD FACSDiva Software v8.0.3</td><td>BD Biosciences</td><td>-</td><td>Adicionado dos Métodos Suplementares</td></tr><tr><td>BD LSRFortessa X-20 cell analyzer</td><td>BD Biosciences</td><td>-</td><td>Entrada existente padronizada para corresponder ao arquivo suplementar</td></tr><tr><td>BioRender</td><td>BioRender</td><td>-</td><td>Usado para criar o fluxograma da Figura 1</td></tr><tr><td>Blotting-grade blocker</td><td>BioRad</td><td>170-6404</td><td></td></tr><tr><td>BODIPY 493/503</td><td>Invitrogen</td><td>D3922</td><td></td></tr><tr><td>Bovine Serum Albumin (BSA)</td><td>Sigma</td><td>A1470</td><td></td></tr><tr><td>CaCl2</td><td>Calbiochem</td><td>208291</td><td></td></tr><tr><td>cDNA Reverse Transcription Kit</td><td>Invitrogen</td><td>4374966</td><td></td></tr><tr><td>Cell Strainer 100 &mu;m, nylon</td><td>Falcon</td><td>352360</td><td></td></tr><tr><td>Cell Strainer 40 &mu;m, nylon</td><td>Falcon</td><td>352340</td><td></td></tr><tr><td>Collagenase type II</td><td>Gibco</td><td>17101-015</td><td></td></tr><tr><td>Dexamethasone</td><td>Sigma</td><td>D4902</td><td></td></tr><tr><td>DMEM Powder, High Glucose, Pyruvate</td><td>Thermo Scientific</td><td>12800017</td><td></td></tr><tr><td>DMEM/F-12, no phenol red</td><td>Thermo Scientific</td><td>21041025</td><td></td></tr><tr><td>DMEM/F-12, powder</td><td>Gibco</td><td>12500062</td><td></td></tr><tr><td>DMSO</td><td>Sigma</td><td>D8418</td><td>Solvente para o meio de indução; fornecedor/catálogo não fornecido</td></tr><tr><td>EMBed-812 EMBEDDING KIT (Epon)</td><td>Electron Microscopy Sciences</td><td>14120</td><td></td></tr><tr><td>Ethanol absolute</td><td>Merck</td><td>100983</td><td></td></tr><tr><td>Ethylene glycol-bis(&beta;-aminoethyl ether)-N,N,N&prime;,N&prime;-tetraacetic acid tetrasodium salt (EGTA)</td><td>Merck</td><td>324626</td><td></td></tr><tr><td>FAST SYBR MASTER MIX</td><td>Thermo Scientific</td><td>4385612</td><td></td></tr><tr><td>FCCP</td><td>Sigma</td><td>C2920</td><td></td></tr><tr><td>Fetal bovine serum</td><td>Capricorn Scientific</td><td>BS-16A</td><td></td></tr><tr><td>Floreada.io</td><td>Floreada.io</td><td>-</td><td>Software de análise de citometria de fluxo</td></tr><tr><td>Fluoromount-G</td><td>Thermo Scientific</td><td>7984-25</td><td></td></tr><tr><td>GAPDH Antibody</td><td>Santa Cruz</td><td>sc-32233</td><td></td></tr><tr><td>Glucose</td><td>Gibco</td><td>15023-021</td><td></td></tr><tr><td>Glutamate</td><td>Sigma</td><td>G1626</td><td></td></tr><tr><td>Glutaraldehyde 25% Aqueous Solution</td><td>Electron Microscopy Sciences</td><td>16210</td><td></td></tr><tr><td>Goat anti-Mouse IgG (H+L) Secondary Antibody, HRP</td><td>Invitrogen</td><td>62-6520</td><td></td></tr><tr><td>GraphPad Prism v10.6.1</td><td>GraphPad Software</td><td>-</td><td>Software de análise estatística/regressão logarítmica</td></tr><tr><td>Halt Protease and Phosphatase Inhibitor Cocktail 100X</td><td>Thermo Scientific</td><td>78442</td><td></td></tr><tr><td>HEPES</td><td>Biological Industries</td><td>41-122-100</td><td></td></tr><tr><td>Histone H3 (1G1)</td><td>Santa Cruz</td><td>sc-517576</td><td></td></tr><tr><td>Hoechst 33342</td><td>Life Technologies</td><td>H3570</td><td></td></tr><tr><td>ImageJ v1.54s</td><td>NIH</td><td>-</td><td>Software de análise de imagem de gotículas de lipídio</td></tr><tr><td>Immun-Blot PVDF Membrane</td><td>BioRad</td><td>1620177</td><td></td></tr><tr><td>Indomethacin</td><td>Sigma</td><td>I7378</td><td></td></tr><tr><td>Insulin syringe</td><td>Cranberry</td><td>AAJECIN3</td><td>Usado para homogeneização celular; fornecedor/catálogo não fornecido</td></tr><tr><td>KCl</td><td>Calbiochem</td><td>529552</td><td></td></tr><tr><td>KH2PO4</td><td>Calbiochem</td><td>529568</td><td></td></tr><tr><td>KHCO3</td><td>Sigma</td><td>60339</td><td></td></tr><tr><td>LAMP-1 Antibody</td><td>Abcam</td><td>ab25245</td><td></td></tr><tr><td>Lane Marker Reducing Sample Buffer</td><td>Thermo Scientific</td><td>39000</td><td></td></tr><tr><td>Malate</td><td>Sigma</td><td>M1000</td><td></td></tr><tr><td>MgCl2</td><td>Calbiochem</td><td>208291</td><td>Componente da solução de ensaio mitocondrial

transferência mitocondrial exógena em adipócitos marrons diferenciadores e preadipócitos deficientes em agpat2

Exogenous Mitochondrial Transfer in Differentiating Brown Adipocytes and AGPAT2-Deficient Preadipocytes

Transferência mitocondrial exógena em adipócitos marrons diferenciadores e preadipócitos deficientes em AGPAT2

Mitochondria are key signaling hubs; however, whether mitochondrial mass expansion is mechanistically required for differentiation remains an open ...

exogenous-mitochondrial-transfer-differentiating-brown-adipocytes

Research

JoVE Journal

Biology

Biologia

Genética induzível Destino Mapping (GIFM) marcas e faixas de células com o controle espacial e temporal...

Uma Abordagem Prática para mapeamento genético induzível Fate: Um Guia Visual para Mark e Track Células In Vivo</em

Visualizando Localização RNA em Xenopus Oócitos

Aqui relatamos protocolos para detectar proteínas endógenas e exógenas de centrômero-cinetócoro em células humanas e quantificar esses níveis de proteína em centrômeros-cinetocoros por coloração imunofluorescente indireta através do uso de fixação (fixação de paraformaldeído, acetona ou metanol).

Proteínas Análise de imunofluorescência de endógenos e exógenos Centromere-cinetócoro

A análise da relação estrutura-função mitocondrial é necessária para uma compreensão completa dos mecanismos regulatórios da funcionalidade mitocondrial. Métodos específicos para estudar a estrutura e função mitocondrial em ovários vivos e fixos de Drosophila são descritos e demonstrados neste artigo.

Estudar mitocondrial estrutura e função em Drosophila ovários

As mitocôndrias pode utilizar o potencial electroquímico através da membrana seu interior (Δ Ψm) para sequestrar cálcio (Ca2 +), o que lhes permite moldar citosólica de Ca 2+ de sinalização dentro da célula. Descreve-se um método para medir simultaneamente mitocôndrias Ca 2+ de fixação e ΔΨ m em células vivas utilizando corantes fluorescentes e microscopia confocal.

Medição simultânea de mitocondrial de cálcio e mitocondrial potencial de membrana em células vivas por Microscopia fluorescente

Aqui, apresentamos um protocolo para a isolação do RNA, DNA e proteínas da mesma amostra, em um esforço para reduzir a variação, melhorar a reprodutibilidade e facilitar interpretações.

Combinado nucleotídeos e proteínas extrações em Caenorhabditis elegans

Este relatório descreve um protocolo para o isolamento simultâneo de preadipócitos primários marrons e brancos de camundongos recém-nascidos. Células isoladas podem ser cultivadas na cultura e induzidas a se diferenciar em adipócitos brancos e marrons totalmente maduros. O método permite a caracterização genética, molecular e funcional das células de gordura primárias na cultura.

Isolamento e diferenciação de preadipócitos brancos e marrons primários de camundongos recém-nascidos

Neste trabalho, descrevemos um protocolo modificado para testar o flux de substrato respiratório mitocondrial usando perfringolysina recombinante O em combinação com respirometria à base de microplato. Com este protocolo, mostramos como a metformina afeta a respiração mitocondrial de duas linhas diferentes de células tumorais.

Medindo flux de substrato mitocondrial em células o-permeabilizadas de permeabilized recombinante

Este manuscrito descreve um protocolo para medir a taxa metabólica basal e a capacidade oxidativa de adipócitos termogênicos em camundongos obesos.

Determinando o gasto energético basal e a capacidade de adipócitos termogênicos para gastar energia em camundongos obesos

Este protocolo descreve o isolamento de pré-adipócitos de camundongos da gordura subcutânea, sua diferenciação em adipócitos maduros e a indução de resistência à insulina. A ação da insulina é avaliada pela fosforilação/ativação de membros da via de sinalização da insulina através do western blot. Este método permite a determinação direta da resistência/sensibilidade à insulina em adipócitos primários.

Adipócitos diferenciados de camundongos em cultura primária: um modelo de resistência à insulina

Pontificia Universidad Católica de Chile.  Mitocôndrias exógenas de origem humana e murinha podem ser internalizadas diferenciando preadipócitos e acompanhadas ao longo da diferenciação adipogênica usando ferramentas de imagem e genéticas. Embora essas mitocôndrias interajam fisicamente com a rede mitocondrial endógena, elas não aumentam a expressão dos marcadores associados aos adipócitos marrons maduros.

Transferência mitocondrial exógena em adipócitos marrons diferenciadores e preadipócitos deficientes em AGPAT2

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