brown preadipocytes  (ATCC)

Journal: bioRxiv

Article Title: Orai1-mediated Ca 2+ Entry Regulates Lipolysis and Mitochondrial Activation in Brown Adipose Thermogenesis

doi: 10.64898/2026.04.24.718619

Figure Lengend Snippet: a Model illustrating that cold-induced Ca 2+ influx via Orai1 is essential for mitochondrial Ca 2+ uptake and subsequent thermogenesis. b Rhod-2 fluorescence imaging of mitochondrial Ca 2+ in siCtrl and siOrai1 cells. c Quantification of mitochondrial Ca 2+ uptake following temperature shifts (37 °C to 15 °C). d Transmission electron microscopy images of BAT from 6 hr cold-exposed control and Orai1 BKO mice. e Western blot analysis of OXPHOS proteins in control and Orai1 BKO BAT after cold exposure (4 °C) for 6 hrs. f Western blot analysis of OXPHOS proteins during differentiation of preadipocytes to brown adipocytes treated with siOrai1. g Mitochondrial DNA copy number measured in control and Orai1 -deficient cells with CL-316,243 treatment. h OCR analysis of Orai1 -deficient adipocytes treated with CL-316,243. i FAO assay of Orai1 -deficient adipocytes treated with CL-316,243. Data are presented as mean ± SEM. Statistical significance: * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001.

Article Snippet: Fully differentiated adipocytes (day 8) were used for in vitro experiments. hTERT-immortalized human brown preadipocytes (ATCC-CRL-4062) were maintained in DMEM/F12 medium supplemented with 10 % FBS.

Techniques: Fluorescence, Imaging, Transmission Assay, Electron Microscopy, Control, Western Blot

Journal: Molecules

Article Title: Effects of Resveratrol on Adipocytes: Evidence from In Vitro and In Vivo Studies

doi: 10.3390/molecules29225359

Figure Lengend Snippet: Summary of the effects of resveratrol and metabolites on adipocytes in cell culture (in vitro).

Article Snippet: Ku et al. [ ] examined the effects of RSV on primary brown preadipocytes isolated from the interscapular brown adipose tissue (iBAT) of 4-week-old chow-fed Sprague Dawley rats.

Techniques: Cell Culture, Activity Assay

Journal: Molecules

Article Title: Effects of Resveratrol on Adipocytes: Evidence from In Vitro and In Vivo Studies

doi: 10.3390/molecules29225359

Figure Lengend Snippet: Summary of the effects of resveratrol and metabolites on adipocytes in cell culture (in vitro).

Article Snippet: [ ] (2016) , iBAT brown preadipocytes from Male Sprague Dawley rats , RSV 10, 50, and 100 μM During Differentiation , ↑ UCP-1, p-AMPK, and ER-α Protein Content.

Techniques: Cell Culture, Activity Assay

Journal: Molecules

Article Title: Effects of Resveratrol on Adipocytes: Evidence from In Vitro and In Vivo Studies

doi: 10.3390/molecules29225359

Figure Lengend Snippet: Summary of the effects of resveratrol and metabolites on adipose tissue (in vivo).

Article Snippet: [ ] (2016) , iBAT brown preadipocytes from Male Sprague Dawley rats , RSV 10, 50, and 100 μM During Differentiation , ↑ UCP-1, p-AMPK, and ER-α Protein Content.

Techniques: Clinical Proteomics, Mouse Assay, Fat, Injection, Transplantation Assay, Activity Assay, Isolation

Journal: Nature Communications

Article Title: Hepatic SerpinA1 improves energy and glucose metabolism through regulation of preadipocyte proliferation and UCP1 expression

doi: 10.1038/s41467-024-53835-9

Figure Lengend Snippet: a Workflow of the proteomics analysis of serum from control and Ai-DKO mice at 3 days after tamoxifen injection ( n = 3) to identify factor(s) leading to preadipocyte proliferation. b Results of proteomics analysis performed according to the workflow in ( a ) and identification of SerpinA1. Statistical analysis was performed using two-tailed Student’s t test. p < 0.05, with an absolute |Log 2 (fold change Ai-DKO/Control)| > 0.3. c Serum SerpinA1 levels in control, Ai-IGF1RKO, Ai-IRKO and Ai-DKO mice 3 days after tamoxifen injection. The data are presented as the mean ± SEM ( n = 10, one-way ANOVA post hoc Bonferroni test, *** p < 0.0001). d SerpinA1 mRNA expression in various organs and tissues of 4-month-old C57BL/6 mice with a primer that recognizes all five SerpinA1 paralogs. The data are presented as the mean ± SEM ( n = 3). e Schematic of the EdU assay for the analysis of preadipocyte proliferation after SerpinA1 treatment for 18 h. f Representative images of EdU-stained brown preadipocytes (WT-1 cells) treated with different concentrations of SerpinA1 (0, 50, or 100 µg/mL). Untreated cells served as the negative Controls. The cells with red fluorescence are in S phase of mitosis, and all cells exhibit blue fluorescence. Scale bar = 100 μm. g Quantification of EdU-positive brown preadipocytes (WT-1 cells). The data are presented as the mean ± SEM ( n = 8 technical replicates/group, one-way ANOVA post hoc Bonferroni test, *** p < 0.0001). h : Representative images of EdU-stained white preadipocytes (A41 hWAT-SVF cells) treated with different concentrations of SerpinA1 (0, 100, or 200 µg/mL). Scale bar = 100 μm. i Quantification of EdU-positive white preadipocytes (A41 hWAT-SVF cells). The data are presented as the mean ± SEM ( n = 7 technical replicates/group, two-tailed Student’s t test with Bonferroni’s correction, * p = 0.0101 and ** p = 0.0015). Source data are provided as a Source Data file.

Article Snippet: The immortalized murine brown preadipocytes (WT-1) have been deposited to Millipore Sigma and were authenticated by Millipore Sigma (#SCC255).

Techniques: Control, Injection, Two Tailed Test, Expressing, EdU Assay, Staining, Fluorescence

Journal: Nature Communications

Article Title: Hepatic SerpinA1 improves energy and glucose metabolism through regulation of preadipocyte proliferation and UCP1 expression

doi: 10.1038/s41467-024-53835-9

Figure Lengend Snippet: a Schematic of the protocol used to induce brown preadipocytes to differentiate into mature brown adipocytes. b Relative mRNA expression of genes in mature brown adipocytes treated with 0 or 200 µg/mL SerpinA1 for 6 h ( n = 8 technical replicates/group, *** p < 0.0001). c UCP1 protein expression in mature brown adipocytes treated with 0 or 300 µg/mL SerpinA1 for 24 h ( n = 5). d Quantification of UCP1 protein levels in ( c ), expressed relative to the level of the protein standard ( n = 5 technical replicates/group, ** p = 0.0025). e Representative traces of the oxygen consumption rate (OCR) in mature brown adipocytes treated with 0 or 300 µg/mL SerpinA1 for 16 h (Control n = 9, SerpinA1 n = 10, technical replicates/group, * p < 0.05, ** p < 0.01 and *** p < 0.001). f Quantitation of basal respiration, ATP production (* p = 0.0141), maximal respiration (* p = 0.0185) and spare respiratory capacity (Control n = 9, SerpinA1 n = 10, technical replicates/group). g Schematic of the protocol used to induce white preadipocytes (A41 hWAT-SVF cells) to differentiate into mature white adipocytes. h Relative mRNA expression of genes in mature white adipocytes treated with different concentrations of SerpinA1 (0, 200, or 500 µg/mL) for 16 h (0 µg/mL n = 8, 200 µg/mL n = 8, and 500 µg/mL n = 7, technical replicates/group, * p = 0.0444 and ** p = 0.0017). i Representative traces of the OCR in mature white adipocytes treated with 0 or 500 µg/mL SerpinA1 for 36 h (Control n = 19, SerpinA1 n = 20, technical replicates/group, * p < 0.05, ** p < 0.01 and *** p < 0.001). j Quantitation of basal respiration, ATP production (* p = 0.0132), maximal respiration (*** p < 0.0001) and spare respiratory capacity (*** p < 0.0001) (Control n = 19, SerpinA1 n = 20, technical replicates/group). Data are presented as mean ± SEM. P values were determined using two-tailed Student’s t test: ( b , d – f , i – j ); one-way ANOVA post hoc Bonferroni test: ( h ). Source data are provided as a Source Data file.

Article Snippet: The immortalized murine brown preadipocytes (WT-1) have been deposited to Millipore Sigma and were authenticated by Millipore Sigma (#SCC255).

Techniques: Expressing, Control, Quantitation Assay, Two Tailed Test

Journal: Nature Communications

Article Title: Hepatic SerpinA1 improves energy and glucose metabolism through regulation of preadipocyte proliferation and UCP1 expression

doi: 10.1038/s41467-024-53835-9

Figure Lengend Snippet: a Schematic of the protocol used to transduce brown preadipocytes with adenovirus carrying 3×Flag-tagged SerpinA1. b Adenovirus-mediated 3×Flag-tagged SerpinA1 protein overexpression brown preadipocytes ( n = 3). c Quantification of protein levels in ( b ) ( n = 3, *** p < 0.0001). d Schematic of the protocol used to induce the differentiation of brown preadipocytes infected with adenovirus carrying 3×Flag-tagged SerpinA1. e Relative UCP1 mRNA expression in mature control and SerpinA1-OE brown adipocytes ( n = 12, *** p = 0.0004). f UCP1 protein expression in mature control and SerpinA1-OE brown adipocytes ( n = 3). g Quantification of protein levels in ( f ) ( n = 3, * p = 0.0200). h Representative immunoblots before and after IP ( n = 1) in control and SerpinA1-OE brown preadipocytes. i Potential SerpinA1-interacting proteins identified by mass spectrometry-based proteomics analysis followed by IP. j Identification of EphB2 as a member of the SerpinA1 complex via the coprecipitation of SerpinA1 and EphB2 by immunoblotting ( n = 1). k EphB2 protein expression in control and CRISPR‒Cas9-mediated EphB2-KO brown preadipocytes ( n = 6). l Quantification of protein levels in ( k ) ( n = 6, *** p < 0.0001). m Representative images of EdU-stained control and EphB2-KO brown preadipocytes treated with different concentrations of SerpinA1 (0 or 100 µg/mL, 18 h). Scale bar = 100 μm. n Quantification of EdU-positive control and EphB2-KO brown preadipocytes treated with different concentrations of SerpinA1 (0 or 100 µg/mL, 18 h) (n = 8, *** p < 0.0001). o The protein levels of p-ERK1/2 and T-ERK1/2 in control and EphB2-KO brown preadipocytes treated with different concentrations of SerpinA1 (0 or 300 µg/mL, 24 h) ( n = 3). p Quantification of protein levels in ( o ) ( n = 3, * p = 0.0357 and *** p = 0.0001). Data are presented as mean ± SEM. P values were determined using two-tailed Student’s t test: ( c , e , g , l ); one-way ANOVA post hoc Bonferroni test: ( n , p ). All replicates performed in Fig. 3 were biologically independent cell clones/group. Source data are provided as a Source Data file.

Article Snippet: The immortalized murine brown preadipocytes (WT-1) have been deposited to Millipore Sigma and were authenticated by Millipore Sigma (#SCC255).

Techniques: Transduction, Over Expression, Infection, Expressing, Control, Western Blot, Mass Spectrometry, Staining, Positive Control, Two Tailed Test, Clone Assay