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( a ) Experimental design for beige precursor quantification prior to cold exposure. ( b ) Gating strategy for the identification of beige precursors (PDGFRα+; <t>EBF2+)</t> in the SVF of EAT and IAT from CTR and SPX mice after 3 days of SPX treatment (S). The cells studied were selected, eliminating the immune cells, dead cells, and debris according to size/volume (SSC-H) and complexity (FSC-H), resulting in a reduced SVF. We first identified PDGFRα+ cells (PDGFRα+ PE/Cy7). Then, within PDGFRα+, we selected EBF2+ cells (Ebf2 AlexaF 647). ( c ) Quantification of the abundance (%) of beige precursors (PDGFRα+; EBF2+) in the SVF of EAT and IAT from CTR and SPX mice. n = 4, mean ± SEM. The effects were evaluated by t -test. The value was statistically significant when p < 0.05.
Intracellular Antibody Anti Ebf2 Alexa Fluor 647, supplied by Bioss, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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( a ) Experimental design for beige precursor quantification prior to cold exposure. ( b ) Gating strategy for the identification of beige precursors (PDGFRα+; <t>EBF2+)</t> in the SVF of EAT and IAT from CTR and SPX mice after 3 days of SPX treatment (S). The cells studied were selected, eliminating the immune cells, dead cells, and debris according to size/volume (SSC-H) and complexity (FSC-H), resulting in a reduced SVF. We first identified PDGFRα+ cells (PDGFRα+ PE/Cy7). Then, within PDGFRα+, we selected EBF2+ cells (Ebf2 AlexaF 647). ( c ) Quantification of the abundance (%) of beige precursors (PDGFRα+; EBF2+) in the SVF of EAT and IAT from CTR and SPX mice. n = 4, mean ± SEM. The effects were evaluated by t -test. The value was statistically significant when p < 0.05.
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( a ) Experimental design for beige precursor quantification prior to cold exposure. ( b ) Gating strategy for the identification of beige precursors (PDGFRα+; <t>EBF2+)</t> in the SVF of EAT and IAT from CTR and SPX mice after 3 days of SPX treatment (S). The cells studied were selected, eliminating the immune cells, dead cells, and debris according to size/volume (SSC-H) and complexity (FSC-H), resulting in a reduced SVF. We first identified PDGFRα+ cells (PDGFRα+ PE/Cy7). Then, within PDGFRα+, we selected EBF2+ cells (Ebf2 AlexaF 647). ( c ) Quantification of the abundance (%) of beige precursors (PDGFRα+; EBF2+) in the SVF of EAT and IAT from CTR and SPX mice. n = 4, mean ± SEM. The effects were evaluated by t -test. The value was statistically significant when p < 0.05.
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( a ) Experimental design for beige precursor quantification prior to cold exposure. ( b ) Gating strategy for the identification of beige precursors (PDGFRα+; EBF2+) in the SVF of EAT and IAT from CTR and SPX mice after 3 days of SPX treatment (S). The cells studied were selected, eliminating the immune cells, dead cells, and debris according to size/volume (SSC-H) and complexity (FSC-H), resulting in a reduced SVF. We first identified PDGFRα+ cells (PDGFRα+ PE/Cy7). Then, within PDGFRα+, we selected EBF2+ cells (Ebf2 AlexaF 647). ( c ) Quantification of the abundance (%) of beige precursors (PDGFRα+; EBF2+) in the SVF of EAT and IAT from CTR and SPX mice. n = 4, mean ± SEM. The effects were evaluated by t -test. The value was statistically significant when p < 0.05.

Journal: International Journal of Molecular Sciences

Article Title: Role of Spexin in White Adipose Tissue Thermogenesis under Basal and Cold-Stimulated Conditions

doi: 10.3390/ijms25031767

Figure Lengend Snippet: ( a ) Experimental design for beige precursor quantification prior to cold exposure. ( b ) Gating strategy for the identification of beige precursors (PDGFRα+; EBF2+) in the SVF of EAT and IAT from CTR and SPX mice after 3 days of SPX treatment (S). The cells studied were selected, eliminating the immune cells, dead cells, and debris according to size/volume (SSC-H) and complexity (FSC-H), resulting in a reduced SVF. We first identified PDGFRα+ cells (PDGFRα+ PE/Cy7). Then, within PDGFRα+, we selected EBF2+ cells (Ebf2 AlexaF 647). ( c ) Quantification of the abundance (%) of beige precursors (PDGFRα+; EBF2+) in the SVF of EAT and IAT from CTR and SPX mice. n = 4, mean ± SEM. The effects were evaluated by t -test. The value was statistically significant when p < 0.05.

Article Snippet: Finally, the cells were incubated with the intracellular antibody anti-Ebf2-Alexa Fluor 647 (1:400, Bioss antibodies, Woburn, MA, USA) and washed once.

Techniques:

KEY RESOURCES TABLE

Journal: Cell reports

Article Title: Defective brown adipose tissue thermogenesis and impaired glucose metabolism in mice lacking Letmd1

doi: 10.1016/j.celrep.2021.110104

Figure Lengend Snippet: KEY RESOURCES TABLE

Article Snippet: Primary antibodies used in this study were anti-Letmd1 (LSBio; LS-C335200), anti-Brg1 (Bethyl; A300-813A-T or Cell Signaling; 49360S), anti-Ucp1 (R&D systems; MAB6158), anti-Prdm16 (R&D systems; AF6295), anti-Ebf2 (R&D systems; AF7006), anti-OXPHOS (MitoSciences; MS604), anti-lamin (Santa Cruz Biotechnology; sc-376248), anti-VDAC (Cell Signaling; 4661T), anti-FABP4 (Santa Cruz; sc-271529), anti-Flag (GenScript; A00187), anti-V5 (ThermoFisher; R96025), anti-β-actin (Santa Cruz; sc-47778), anti-Gapdh (Santa Cruz; sc-25778), and anti-vinculin (Sigma, V9131).

Techniques: Recombinant, Cell Culture, Reverse Transcription, SYBR Green Assay, Chromatin Immunoprecipitation, Plasmid Preparation, Software