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Molecular mechanisms of SIRT1 regulation in epididymal adipose tissue. (A) Immunofluorescence co‐localization of SIRT1 (green) and <t>PPARγ</t> (red) with nuclear DAPI staining (blue) (400×). (B) WB analysis of PPARγ, NF‐κB, and HIF1a protein expression. (C) mRNA expression levels of PPARγ, NF‐κB, and HIF1a in epididymal adipose tissue. (D) Validation of DNMT3a overexpression and its effect on SIRT1 protein levels in 3T3‐L1 adipocytes. (E) ChIP analysis of DNMT3a binding to the SIRT1 promoter region. p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001.
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Molecular mechanisms of SIRT1 regulation in epididymal adipose tissue. (A) Immunofluorescence co‐localization of SIRT1 (green) and <t>PPARγ</t> (red) with nuclear DAPI staining (blue) (400×). (B) WB analysis of PPARγ, NF‐κB, and HIF1a protein expression. (C) mRNA expression levels of PPARγ, NF‐κB, and HIF1a in epididymal adipose tissue. (D) Validation of DNMT3a overexpression and its effect on SIRT1 protein levels in 3T3‐L1 adipocytes. (E) ChIP analysis of DNMT3a binding to the SIRT1 promoter region. p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001.
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Molecular mechanisms of SIRT1 regulation in epididymal adipose tissue. (A) Immunofluorescence co‐localization of SIRT1 (green) and PPARγ (red) with nuclear DAPI staining (blue) (400×). (B) WB analysis of PPARγ, NF‐κB, and HIF1a protein expression. (C) mRNA expression levels of PPARγ, NF‐κB, and HIF1a in epididymal adipose tissue. (D) Validation of DNMT3a overexpression and its effect on SIRT1 protein levels in 3T3‐L1 adipocytes. (E) ChIP analysis of DNMT3a binding to the SIRT1 promoter region. p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001.

Journal: Food Science & Nutrition

Article Title: Flavonoids in Rosa roxburghii Tratt Fermentation Broth Ameliorate Obesity via DNMT3a / SIRT1 ‐Mediated Epigenetic Modulation

doi: 10.1002/fsn3.70892

Figure Lengend Snippet: Molecular mechanisms of SIRT1 regulation in epididymal adipose tissue. (A) Immunofluorescence co‐localization of SIRT1 (green) and PPARγ (red) with nuclear DAPI staining (blue) (400×). (B) WB analysis of PPARγ, NF‐κB, and HIF1a protein expression. (C) mRNA expression levels of PPARγ, NF‐κB, and HIF1a in epididymal adipose tissue. (D) Validation of DNMT3a overexpression and its effect on SIRT1 protein levels in 3T3‐L1 adipocytes. (E) ChIP analysis of DNMT3a binding to the SIRT1 promoter region. p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001.

Article Snippet: Immunofluorescence double staining was carried out on the paraffin‐embedded epididymal adipose tissue sections using antibodies against silent information regulator 1 (SIRT1) and peroxisome proliferator‐activated receptor gamma (PPARγ) antibodies (13161‐1‐AP and 16643‐1‐AP; Proteintech, Wuhan, China).

Techniques: Immunofluorescence, Staining, Expressing, Biomarker Discovery, Over Expression, Binding Assay