Journal: Journal of Lipid Research

Article Title: SDCCAG3 inhibits adipocyte hypertrophy and improves obesity-related metabolic disorders via SDCCAG3/SMURF1/PPARγ axis

doi: 10.1016/j.jlr.2025.100772

Figure Lengend Snippet: Determining SDCCAG3 as a regulator of adipocyte metabolism. A: Protein expression levels of SDCCAG3 in IngWAT of mice on NCD or HFD. B: Protein expression levels of SDCCAG3 in EpiWAT of mice on NCD or HFD. C: SDCCAG3 mRNA levels in IngWAT and EpiWAT of mice on NCD or HFD. D and E: Changes in SDCCAG3 protein and mRNA levels during the differentiation of 3T3-L1 cells into mature adipocytes. F: Protein and mRNA levels of SDCCAG3 in differentiated 3T3-L1 cells cultured under normal or high-fat conditions. G: Protein expression levels of PPARγ, C/EBPα and SDCCAG3 with knockdown or overexpression of SDCCAG3 on differentiated 3T3-L1 cells cultured under high-fat conditions. H: SDCCAG3, PPARγ, C/EBPα, FABP4, LPL and ADIPOQ mRNA levels with knockdown or overexpression of SDCCAG3 on differentiated 3T3-L1 cells cultured under high-fat conditions. I and K: Oil red O staining of differentiated 3T3-L1 cells and relative area with knockdown or overexpression of SDCCAG3, scale bar = 100 μm. J and L: BODIPY staining of differentiated 3T3-L1 cells and relative intensity with knockdown or overexpression of SDCCAG3, scale bar = 75 μm. Data were presented as mean ± standard deviation (S.D.). ∗ P < 0.05, ∗∗ P < 0.01, ∗∗∗ P < 0.001, ∗∗∗∗ P < 0.0001.

Article Snippet: For BODIPY staining, cells were first fixed in a 4% paraformaldehyde solution for 30 min and stained with 0.5 μM BODIPY 493/503 (GC42959, GLPBIO) for 30 min, followed by DAPI (C0065, Solarbio) staining for 5 min.

Techniques: Expressing, Cell Culture, Knockdown, Over Expression, Staining, Standard Deviation

Journal: Journal of Lipid Research

Article Title: SDCCAG3 inhibits adipocyte hypertrophy and improves obesity-related metabolic disorders via SDCCAG3/SMURF1/PPARγ axis

doi: 10.1016/j.jlr.2025.100772

Figure Lengend Snippet: SMURF1 mediated the regulation of SDCCAG3 on PPARγ protein stability and adipocyte. A: Co-immunoprecipitation of SDCCAG3 and SMURF1 in 3T3-L1 cells. B: Co-immunoprecipitation of SDCCAG3 and SMURF1 in HEK293T cells exogenously expressing HA-Sdccag3 and Smurf1. C: The mRNA levels of SMURF1 with knockdown or overexpression of SDCCAG3 on differentiated 3T3-L1 cells. D: Protein expression levels of SMURF1 and SDCCAG3 with knockdown or overexpression of SDCCAG3 on differentiated 3T3-L1 cells. E: The HEK293T cells overexpressing Flag-Pparγ were co-transfected with HA-Sdccag3 or Smurf1, and the protein expression levels of Flag-PPARγ, SMURF1 and SDCCAG3 were detected after CHX treatment for 0, 2, 4, and 6h. F: HA-Sdccag3 or Smurf1 were co-transfected with UB and Flag-Pparγ plasmids into HEK293T cells. Flag-PPARγ protein was pulled down by magnetic beads, and the ubiquitination levels were detected by Western blot. G: Protein expression levels of PPARγ, C/EBPα, SDCCAG3 and SMURF1 with co-transfection of Smurf1 and HA-Sdccag3 on differentiated 3T3-L1 cells cultured under high-fat conditions. H: PPARγ, C/EBPα, FABP4, LPL and ADIPOQ mRNA levels with co-transfection of Smurf1 and HA-Sdccag3 on differentiated 3T3-L1 cells cultured under high-fat conditions. I: Oil red O staining and relative area of differentiated 3T3-L1 cells cultured under high-fat conditions with co-transfection of Smurf1 and HA-Sdccag3, scale bar = 100 μm. J: BODIPY staining and relative intensity of differentiated 3T3-L1 cells cultured under high-fat conditions with co-transfection of Smurf1 and HA-Sdccag3, scale bar = 200 μm. Data were presented as mean ± standard deviation (S.D.). ∗ P < 0.05, ∗∗ P < 0.01, ∗∗∗ P < 0.001, ∗∗∗∗ P < 0.0001.

Article Snippet: For BODIPY staining, cells were first fixed in a 4% paraformaldehyde solution for 30 min and stained with 0.5 μM BODIPY 493/503 (GC42959, GLPBIO) for 30 min, followed by DAPI (C0065, Solarbio) staining for 5 min.

Techniques: Immunoprecipitation, Expressing, Knockdown, Over Expression, Transfection, Magnetic Beads, Ubiquitin Proteomics, Western Blot, Cotransfection, Cell Culture, Staining, Standard Deviation

Journal: Journal of Lipid Research

Article Title: SDCCAG3 inhibits adipocyte hypertrophy and improves obesity-related metabolic disorders via SDCCAG3/SMURF1/PPARγ axis

doi: 10.1016/j.jlr.2025.100772

Figure Lengend Snippet: Determining the functional fragment of SDCCAG3 and a SDCCAG3-PPARγ-SDCCAG3 lipid metabolism enhancement loop. A: Protein expression levels of SDCCAG3, PPARγ, C/EBPα in differentiated 3T3-L1 cells transfected with HA-Sdccag3 and treated with GW9662 under high-fat conditions. B: The mRNA levels of SDCCAG3, PPARγ, C/EBPα, FABP4, LPL and ADIPOQ in differentiated 3T3-L1 cells transfected with HA-Sdccag3 and treated with GW9662 under high-fat conditions. C: Differentiated 3T3-L1 cells transfected with HA-Sdccag3 were treated with GW9662 and stained with oil red O, scale bar = 50 μm. D: Differentiated 3T3-L1 cells transfected with HA-Sdccag3 were treated with GW9662 and stained with BODIPY, scale bar = 75 μm. E: Example plots of full-length Sdccag3 and its mutants lacking segments 1–145, 146–261, and 262–432, respectively. F: In HEK293T cells, full-length Sdccag3 and its mutants (mut1, mut2, and mut3) were transfected with Flag-Pparγ, and the co-immunoprecipitation of HA and Flag was detected. G: In HEK293T cells, full-length Sdccag3 and its mutants (mut1, mut2, and mut3) were transfected with Smurf1, and the co-immunoprecipitation of HA and SMURF1 was detected. H: Protein expression levels of PPARγ, C/EBPα and SDCCAG3 with transfection of full-length Sdccag3 and its mutants (mut1, mut2, and mut3) on differentiated 3T3-L1 cells. I: Oil red O staining and relative area of differentiated 3T3-L1 cells with transfection of full-length Sdccag3 and its mutants (mut1, mut2, and mut3), scale bar = 100 μm. J: BODIPY staining and relative intensity of differentiated 3T3-L1 cells with transfection of full-length Sdccag3 and its mutants (mut1, mut2, and mut3), scale bar = 200 μm. K: Luciferase activity was measured after transfection of Sdccag3 or mutant luciferase reporter gene plasmid, renilla luciferase gene plasmid and Pparγ plasmid in HEK293T cells. L: Protein expression levels of PPARγ and SDCCAG3 in differentiated 3T3-L1 cells overexpressing Flag-Pparγ. M: SDCCAG3 mRNA levels in differentiated 3T3-L1 cells overexpressing Flag-Pparγ. Data were presented as mean ± standard deviation (S.D.). ∗ P < 0.05, ∗∗ P < 0.01, ∗∗∗ P < 0.001, ∗∗∗∗ P < 0.0001.

Article Snippet: For BODIPY staining, cells were first fixed in a 4% paraformaldehyde solution for 30 min and stained with 0.5 μM BODIPY 493/503 (GC42959, GLPBIO) for 30 min, followed by DAPI (C0065, Solarbio) staining for 5 min.

Techniques: Functional Assay, Expressing, Transfection, Staining, Immunoprecipitation, Luciferase, Activity Assay, Mutagenesis, Plasmid Preparation, Standard Deviation