Journal: Science Advances

Article Title: Abnormal mitochondrial structure and function in brown adipose tissue of SLC35A4-MP knockout mice

doi: 10.1126/sciadv.ads7381

Figure Lengend Snippet: ( A ) Representative SLC35A4-MP blot in various tissues, including brain, kidney, lung, spleen, testis, BAT, eWAT, skeletal muscle (gastrocnemius), heart, and liver from male C57BL6 wild-type mice. Ponceau staining served as a protein level control. ( B ) HFD and chow diet feeding schematic and protocol for BAT collection were created in BioRender. Saghatelian, A. (2025) https://BioRender.com/9luycjt . ( C ) Representative SLC35A4-MP blot and ( D ) quantification in BAT of HFD-fed C57BL/6 male mice ( n = 4 per group). ( E ) qPCR analysis of Slc35a4 mRNA expression in BAT of HFD-fed C57BL/6 male mice ( n = 4 to 6 per group). Data presented as means ± SEM, with * indicating statistical significance ( P < 0.05, unpaired t test) compared to the chow diet.

Article Snippet: Antigen affinity purified SLC35A4 MP antiserum was covalently coupled to Affi-Gel10 N -hydroxysuccinimide–activated resin per the manufacturer’s instructions (Bio-Rad, catalog no. 1536099), 1.2 mg of antiserum to 1.5 ml of bed volume resin.

Techniques: Staining, Control, Expressing

Journal: Science Advances

Article Title: Abnormal mitochondrial structure and function in brown adipose tissue of SLC35A4-MP knockout mice

doi: 10.1126/sciadv.ads7381

Figure Lengend Snippet: Effects of 12-Week HFD in female SLC35A4-MP KO (KO) and WT mice. ( A ) Initial and 12-week body weight under HFD feeding (two-way ANOVA followed by Šídák’s multiple comparisons test). ( B ) Fat tissue weight; ( C ) fasting blood glucose levels; ( D ) glucose tolerance test (GTT), also expressed as ( E ) the area under the curve (AUC); ( F ) BAT histology: hematoxylin and eosin stain in BAT. Scale bar, 100 μm. ( G ) Quantification of lipid droplet (LD) size in micrometer. ( H to J ) LC-MS/MS–based untargeted lipidomics of BAT from SLC35A4-MP KO mice and their controls after 12 weeks of HFD feeding ( n = 5 per group). (I) The total sum of triacylglycerol species detected in BAT. (J) Heatmap showing the total sum levels of phosphatidic acid (PA, P < 0.001), acylcarnitines (AcCa), phosphatidylinositol (PI), phosphatidylserine (PS), lysophosphatidylcholine (LPC), phosphatidylglycerol (PG), phosphatidylcholine (PC), lysophosphatidylethanolamine (LPE), lysophosphatidylglycerol (LPG, P < 0.02), phosphatidylethanolamine (PE, P < 0.007), and cardiolipin (CL, P < 0.007). Data presented as means ± SEM, with * indicating statistical significance ( *P < 0.05, **** P < 0.0001, unpaired t test) compared to the WT mice. ns, not significant.

Article Snippet: Antigen affinity purified SLC35A4 MP antiserum was covalently coupled to Affi-Gel10 N -hydroxysuccinimide–activated resin per the manufacturer’s instructions (Bio-Rad, catalog no. 1536099), 1.2 mg of antiserum to 1.5 ml of bed volume resin.

Techniques: H&E Stain, Liquid Chromatography with Mass Spectroscopy

Journal: Science Advances

Article Title: Abnormal mitochondrial structure and function in brown adipose tissue of SLC35A4-MP knockout mice

doi: 10.1126/sciadv.ads7381

Figure Lengend Snippet: Twelve-week HFD or chow diet in male SLC35A4-MP KO and WT mice. ( A ) Mitochondrial tomography slices: 1.6-nm thickness slices from the center of EM tomography volumes of BAT mitochondria from BAT of male WT mice (left) and SLC35A4-MP KO (right) mice with increased mitochondrial area and decreased numbers (scale bar, 1 μm). ( B ) Quantification of the mitochondrial profile area (size) ( n = 14 to 54 per group), ( C ) number of mitochondria per unit area of cytoplasm (#mitochondria per square micrometer) ( n = 10 per group), ( D ) mitochondrial shape (ratio major and minor axis) ( n = 14 to 54 per group), and ( E ) cristae density ( n = 10 mito per group). Data presented as means ± SEM, with * indicating statistical significance [* P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001, two-way analysis of variance (ANOVA)] compared to the WT mice.

Article Snippet: Antigen affinity purified SLC35A4 MP antiserum was covalently coupled to Affi-Gel10 N -hydroxysuccinimide–activated resin per the manufacturer’s instructions (Bio-Rad, catalog no. 1536099), 1.2 mg of antiserum to 1.5 ml of bed volume resin.

Techniques: Tomography

Journal: Science Advances

Article Title: Abnormal mitochondrial structure and function in brown adipose tissue of SLC35A4-MP knockout mice

doi: 10.1126/sciadv.ads7381

Figure Lengend Snippet: ( A to D ) TMT-labeled proteomics analysis of BAT from male SLC35A4-MP KO mice fed an HFD for 12 weeks ( n = 4 per group). Proteomic samples were processed using a standard TMT workflow, resulting in high proteome coverage with more than 2000 proteins detected. (A) Levels of mitochondrial metabolic protein Perm1, Chchd7, and Ndufs8 ( n = 4 per group) and (B) relative abundance of inflammation-related proteins in BAT, including galectin-3 (Lgals3), macrophage-capping protein (Capg), tapasin (Tapbp), and intercellular adhesion molecule 1 (Icam1). (C) Representative F4/80 immunohistochemistry images with eosin counterstaining (40× magnification). (D) Quantification of F4/80 + staining area using ImageJ (% area; n = 5 per group). ( E ) qPCR analysis of Cd86 gene expression and ( F ) proinflammatory cytokines in BAT of male and ( G ) female mice ( n = 5 per group). ( H to J ) Respiratory ratio analysis in BMDMs activated with IL-4 from male SLC35A4-MP KO and WT mice fed with chow diet. (I) Basal respiratory capacity (J) ATP-linked respiratory capacity using Seahorse XF96 ( n = 12 per group). ( K to M ) Gene expression of proinflammatory cytokines in BMDMs activated with IFN-γ + LPS or IL-4, (K) Tnf-a mRNA, (L) Il1b mRNA, (M) Il12b mRNA (two-way ANOVA followed by Šídák’s multiple comparisons test, n = 3 per group). Data presented as means ± SEM, with * indicating statistical significance ( *P < 0.05, ** P < 0.01, **** P < 0.0001 unpaired t test for all other panels) compared to the WT mice.

Article Snippet: Antigen affinity purified SLC35A4 MP antiserum was covalently coupled to Affi-Gel10 N -hydroxysuccinimide–activated resin per the manufacturer’s instructions (Bio-Rad, catalog no. 1536099), 1.2 mg of antiserum to 1.5 ml of bed volume resin.

Techniques: Labeling, Immunohistochemistry, Staining, Gene Expression

Journal: Science Advances

Article Title: Abnormal mitochondrial structure and function in brown adipose tissue of SLC35A4-MP knockout mice

doi: 10.1126/sciadv.ads7381

Figure Lengend Snippet: ( A ) Representative SLC35A4-MP blot and ( B ) quantification of SLC35A4-MP levels in immortalized murine interscapular brown preadipocytes during differentiation of mature brown adipocytes by Western blot ( n = 3 per group). ( C ) qPCR analysis of Slc35a4 mRNA expression. ( D ) Representative Western blot and ( E ) quantification of SLC35A4-MP levels in BAT from 12-week-old male WT mice after 3 days of cold exposure (chronic cold exposure) ( n = 3 per group). ( F ) qPCR evaluation of Slc35a4 mRNA expression in BAT following chronic cold exposure ( n = 3 to 4 per group). ( G ) qPCR analysis of brown adipocyte markers, Ucp1 , Ppargc1a , Prdm16 , and Elovl3 mRNA in primary brown adipocytes differentiated from the stroma vascular fraction of BAT derived from 1-month-old male SLC35A4-MP KO and WT mice ( n = 3 per group). ( H ) OCR profile of primary brown adipocytes from SLC35A4-MP KO and WT mice. ( I ) Quantification of basal, ( J ) maximal, and ( K ) ATP-linked respiration ( n = 10 per group). Data presented as means ± SEM, with * indicating statistical significance ( *P < 0.05, ** P < 0.01, **** P < 0.0001, unpaired t test) compared to the WT.

Article Snippet: Antigen affinity purified SLC35A4 MP antiserum was covalently coupled to Affi-Gel10 N -hydroxysuccinimide–activated resin per the manufacturer’s instructions (Bio-Rad, catalog no. 1536099), 1.2 mg of antiserum to 1.5 ml of bed volume resin.

Techniques: Western Blot, Expressing, Derivative Assay

Journal: Science Advances

Article Title: Abnormal mitochondrial structure and function in brown adipose tissue of SLC35A4-MP knockout mice

doi: 10.1126/sciadv.ads7381

Figure Lengend Snippet: Effects of acute and chronic cold exposure on 12-week-old male SLC35A4-MP KO and WT mice fed a chow diet. ( A and B ) Rectal body temperature during cold exposure ( n = 6 per group). ( C ) LC-MS/MS–based untargeted lipidomics of BAT from SLC35A4-MP KO mice and their controls after 3 days of cold exposure ( n = 3 per group). ( D ) Acylcarnitine levels in BAT (two-way ANOVA followed by Šídák’s multiple comparisons test, n = 3 per group). ( E ) Mitochondrial tomography slices: 1.6-nm thickness slices from the center of EM tomography volumes of BAT mitochondria from BAT of WT mice (left) and SLC35A4-MP KO (right) male mice after chronic cold challenge (scale bar, 1 μm). RT, room temperature. ( F ) Quantification of the mitochondria profile area (size in square micrometer) ( n = 21 to 54 per group), ( G ) number of mitochondria per unit area of cytoplasm (#mitochondria per square micrometer) ( n = 10 per group), and ( H ) mitochondrial shape (ratio major and minor axis) ( n = 21 to 54 per group). Data presented as means ± SEM, with * indicating statistical significance ( *P < 0.05, ** P < 0.01, **** P < 0.0001, unpaired t test for all other panels) compared to the WT mice.

Article Snippet: Antigen affinity purified SLC35A4 MP antiserum was covalently coupled to Affi-Gel10 N -hydroxysuccinimide–activated resin per the manufacturer’s instructions (Bio-Rad, catalog no. 1536099), 1.2 mg of antiserum to 1.5 ml of bed volume resin.

Techniques: Liquid Chromatography with Mass Spectroscopy, Tomography

Journal: Science Advances

Article Title: Abnormal mitochondrial structure and function in brown adipose tissue of SLC35A4-MP knockout mice

doi: 10.1126/sciadv.ads7381

Figure Lengend Snippet: A schematic model illustrating how SLC35A4-MP supports mitochondrial structure and lipid metabolism in BAT. Loss of SLC35A4-MP leads to altered mitochondrial morphology, reduced oxidative capacity, acylcarnitine accumulation, and inflammation, ultimately impairing thermogenic function. Created in BioRender. Saghatelian, A. (2025) https://BioRender.com/38j8mtz .

Article Snippet: Antigen affinity purified SLC35A4 MP antiserum was covalently coupled to Affi-Gel10 N -hydroxysuccinimide–activated resin per the manufacturer’s instructions (Bio-Rad, catalog no. 1536099), 1.2 mg of antiserum to 1.5 ml of bed volume resin.

Techniques:

Journal: PLOS Global Public Health

Article Title: Performance evaluation of SARS-CoV-2 rapid diagnostic tests in Nigeria: A cross-sectional study

doi: 10.1371/journal.pgph.0003371

Figure Lengend Snippet: List of COVID-19 rapid diagnostic test kits evaluated.

Article Snippet: 4 , MP Rapid SARS-CoV-2 Antigen Card , MP Biomedicals, Germany , 96.49% (93.11%-99.87%) , 99.07% (98.26%-99.98%) , Nucleocapsid protein.

Techniques: Diagnostic Assay