Journal: Cell Death Discovery

Article Title: Irisin inhibits adipogenic differentiation of bone marrow mesenchymal stem cells through the SIRT1/RANBP2/FTO signaling axis and protects against osteoporosis

doi: 10.1038/s41420-026-02976-5

Figure Lengend Snippet: A The effects of 10 μM, 25 μM, 50 μM, 75 μM, 100 μM, and 125 μM concentrations of irisin on the proliferation of BMMSCs were detected on days 12 h, 24 h, 36 h, and 48 h using the CCK-8 assay. B Flow chart of experiments to detect the osteogenic differentiation of BMMSCs. C Representative images of ARS staining during osteogenic (Osteo) differentiation in BMMSCs, and quantitative analysis of ARS staining. D Representative images of ALP staining during osteogenic (Osteo) differentiation in BMMSCs, and quantitative analysis of ALP staining. E After the addition of irisin, qRT-PCR was performed to analyze the mRNA levels of osteogenic (Osteo) related genes ( Runx2 , Col1 , and BMP2 ) in BMMSCs during the osteogenic differentiation. F Flow chart of experiments to detect the adipogenic differentiation of BMMSCs. G After the addition of irisin, qRT-PCR was performed to analyse the mRNA levels of adipogenic (Adipo) related genes ( Pparγ , C/ebpα , and C/ebpβ ) in BMMSCs during the adipogenic differentiation. H Representative images of Oil red O staining after adding irisin, and quantitative analysis of Oil red O staining. I After the addition of irisin, the levels of adipogenic (Adipo) related protein (PPARγ, C/EBPα, and C/EBPβ) in BMMSCs during the adipogenic differentiation were measured by western blotting. The values are mean ± SD of at least three independent experiments; n.s. p > 0.05, * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001.

Article Snippet: In brief, equal amounts of protein samples were separated on polyacrylamide gels and transferred to nitrocellulose membranes, which were closed for 1 h using 5% skimmed milk powder, and the membranes obtained were incubated with anti-FNDC5 antibody (#23995-1-AP, 1:1000, Proteintech, China), anti-PPARγ antibody (#7273, 1:1000, Santa Cruz Biotechnology, USA), anti-C/EBPα antibody (#2295, 1:1000, Cell Signaling Technology, USA), anti-C/EBPβ antibody (#3087, 1:1000, Cell Signaling Technology, USA), anti-SIRT1 antibody (#8469, 1:1000, Cell Signaling Technology, USA), anti-FTO antibody (#27226-1-AP, Proteintech, China), anti-RANBP2 antibody (#ab315458, 1:1000, Abcam, USA), and anti-GAPDH antibody (#10494-1, 1:1000, proteintech, China) were incubated at 4 °C overnight.

Techniques: CCK-8 Assay, Staining, Quantitative RT-PCR, Western Blot

Journal: Cell Death Discovery

Article Title: Irisin inhibits adipogenic differentiation of bone marrow mesenchymal stem cells through the SIRT1/RANBP2/FTO signaling axis and protects against osteoporosis

doi: 10.1038/s41420-026-02976-5

Figure Lengend Snippet: A Schematic workflow of the RNA-seq. B Volcanic maps of RNA-Seq in the PBS group and irisin group. C Heatmap of mRNA expression in the PBS group and irisin group, with high and low expression levels shown in red and blue respectively. D The mRNA levels of Serpina3n , Nos2 , Rdh9 , Serpina3m , and Sirt1 were detected by qRT-PCR. E qRT-PCR and western blotting confirmation of Sirt1 knockdown BMMSCs. F mRNA expressions of Pparγ , C/ebpα , and C/ebpβ in Sirt1 knockdown cells with or without irisin treatment. G Protein expressions of PPPARγ, C/EBPα, and C/EBPβ in Sirt1 knockdown cells with or without irisin treatment. H Representative images of Oil red O staining in Sirt1 knockdown cells with or without irisin treatment, and quantitative analysis of Oil red O staining. The values are mean ± SD of at least three independent experiments; n.s. p > 0.05, * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001.

Article Snippet: In brief, equal amounts of protein samples were separated on polyacrylamide gels and transferred to nitrocellulose membranes, which were closed for 1 h using 5% skimmed milk powder, and the membranes obtained were incubated with anti-FNDC5 antibody (#23995-1-AP, 1:1000, Proteintech, China), anti-PPARγ antibody (#7273, 1:1000, Santa Cruz Biotechnology, USA), anti-C/EBPα antibody (#2295, 1:1000, Cell Signaling Technology, USA), anti-C/EBPβ antibody (#3087, 1:1000, Cell Signaling Technology, USA), anti-SIRT1 antibody (#8469, 1:1000, Cell Signaling Technology, USA), anti-FTO antibody (#27226-1-AP, Proteintech, China), anti-RANBP2 antibody (#ab315458, 1:1000, Abcam, USA), and anti-GAPDH antibody (#10494-1, 1:1000, proteintech, China) were incubated at 4 °C overnight.

Techniques: RNA Sequencing, Expressing, Quantitative RT-PCR, Western Blot, Knockdown, Staining

Journal: Cell Death Discovery

Article Title: Irisin inhibits adipogenic differentiation of bone marrow mesenchymal stem cells through the SIRT1/RANBP2/FTO signaling axis and protects against osteoporosis

doi: 10.1038/s41420-026-02976-5

Figure Lengend Snippet: A Flowchart of SIRT1 destabilized FTO via RANBP2-mediated SUMOylation. B Fto mRNA expression in Sirt1 knockdown or Sirt1 -overexpressed cells was assayed by qRT-PCR. FTO protein expression in Sirt1 knockdown or Sirt1 -overexpressed cells was assayed by western blotting. C Fto mRNA expression in Sirt1 knockdown cells with or without irisin treatment was assayed by qRT-PCR. FTO protein expression in Sirt1 knockdown cells with or without irisin treatment was assayed by western blotting. D Immunoblotting of adipogenic related proteins (PPARγ, C/EBPα, and C/EBPβ) in Sirt1 -overexpressed cells with or without Fto overexpression. E Representative images of Oil red O staining in Sirt1 -overexpressed cells with or without Fto overexpression, and quantitative analysis of Oil red O staining. The values are mean ± SD of at least three independent experiments; n.s. p > 0.05, * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001.

Article Snippet: In brief, equal amounts of protein samples were separated on polyacrylamide gels and transferred to nitrocellulose membranes, which were closed for 1 h using 5% skimmed milk powder, and the membranes obtained were incubated with anti-FNDC5 antibody (#23995-1-AP, 1:1000, Proteintech, China), anti-PPARγ antibody (#7273, 1:1000, Santa Cruz Biotechnology, USA), anti-C/EBPα antibody (#2295, 1:1000, Cell Signaling Technology, USA), anti-C/EBPβ antibody (#3087, 1:1000, Cell Signaling Technology, USA), anti-SIRT1 antibody (#8469, 1:1000, Cell Signaling Technology, USA), anti-FTO antibody (#27226-1-AP, Proteintech, China), anti-RANBP2 antibody (#ab315458, 1:1000, Abcam, USA), and anti-GAPDH antibody (#10494-1, 1:1000, proteintech, China) were incubated at 4 °C overnight.

Techniques: Expressing, Knockdown, Quantitative RT-PCR, Western Blot, Over Expression, Staining

Journal: Cell Death Discovery

Article Title: Irisin inhibits adipogenic differentiation of bone marrow mesenchymal stem cells through the SIRT1/RANBP2/FTO signaling axis and protects against osteoporosis

doi: 10.1038/s41420-026-02976-5

Figure Lengend Snippet: A RANBP2 protein expression in Sirt1 knockdown cells with or without irisin treatment was assayed by western blotting. B Co-immunoprecipitation of SIRT1 with RANBP2 in BMMSCs with or without irisin treatment. C Colocalization of SIRT1 and RANBP2 after treatment with or without irisin in BMMSCs was exhibited in confocal immunofluorescent images. D Representative images of Oil red O staining in Sirt1 -overexpressed cells with or without Ranbp2 knockdown, and quantitative analysis of Oil red O staining. E mRNA expressions of Pparγ , C/ebpα , and C/ebpβ in Sirt1 -overexpressed cells with or without Ranbp2 knockdown. F Fto mRNA expression in Sirt1 -overexpressed cells with or without Ranbp2 knockdown was assayed by qRT-PCR. G FTO protein expression in Sirt1-overexpressed cells with or without Ranbp2 knockdown was assayed by western blotting. H Co-immunoprecipitation was used to detect the interactions of FTO with SIRT1 and RANBP2 proteins in irisin-treated or non-treated BMMSCs. Co-immunoprecipitation was used to detect the interactions of FTO with SUMO2/3 in irisin-treated or non-treated BMMSCs. The values are mean ± SD of at least three independent experiments; n.s. p > 0.05, * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001.

Article Snippet: In brief, equal amounts of protein samples were separated on polyacrylamide gels and transferred to nitrocellulose membranes, which were closed for 1 h using 5% skimmed milk powder, and the membranes obtained were incubated with anti-FNDC5 antibody (#23995-1-AP, 1:1000, Proteintech, China), anti-PPARγ antibody (#7273, 1:1000, Santa Cruz Biotechnology, USA), anti-C/EBPα antibody (#2295, 1:1000, Cell Signaling Technology, USA), anti-C/EBPβ antibody (#3087, 1:1000, Cell Signaling Technology, USA), anti-SIRT1 antibody (#8469, 1:1000, Cell Signaling Technology, USA), anti-FTO antibody (#27226-1-AP, Proteintech, China), anti-RANBP2 antibody (#ab315458, 1:1000, Abcam, USA), and anti-GAPDH antibody (#10494-1, 1:1000, proteintech, China) were incubated at 4 °C overnight.

Techniques: Expressing, Knockdown, Western Blot, Immunoprecipitation, Staining, Quantitative RT-PCR

Journal: Animal Cells and Systems

Article Title: Quercus infectoria gall extract modulates adipocyte differentiation and lipid metabolism through dual regulation of adipogenesis and lipolysis in 3T3-L1 adipocytes

doi: 10.1080/19768354.2026.2623321

Figure Lengend Snippet: Quercus infectoria gall (QIG) extract inhibits the expression of key adipogenic transcription factors and enzymes in differentiating 3T3-L1 adipocytes in a dose-dependent manner. The differentiation/activation medium (D/A) was added to the 3T3-L1 preadipocytes with or without QIG extract at varying concentrations (3.12, 6.25, 12.5, and 25 μg/mL). The treatment was performed for 3 days before harvesting the cells for Western blot assays. (a) Representative Western blot data set performed using anti-C/EBPβ, anti-C/EBPα, anti-PPARγ, anti-FAS, and anti-β-actin antibodies. (b) Quantification of the levels of C/EBPβ, C/EBPα, PPARγ, and FAS proteins, as shown in (a). Data represent three independent experiments performed in triplicate and are expressed as mean ± SD (error bars). Different letters indicate statistically significant differences ( p < 0.05).

Article Snippet: Primary antibodies used in this study included C/EBPβ (LAP) Antibody (#3087), C/EBPα (D56F10) XP® Rabbit mAb (#8178), PPARγ (81B8) Rabbit mAb (#2443), Fatty Acid Synthase (FAS) (C20G5) Rabbit mAb (#3180), and Phospho-HSL (Ser660) Rabbit mAb (#45804), all purchased from Cell Signaling Technology (Danvers, MA, USA); HSL Rabbit pAb (A15686) and ATGL/PNPLA2 Rabbit pAb (A6245) from Abclonal (Wuhan, China); and β-actin Rabbit mAb (MABT523) from Merck (Darmstadt, Germany).

Techniques: Expressing, Activation Assay, Western Blot

Journal: Animal Cells and Systems

Article Title: Quercus infectoria gall extract modulates adipocyte differentiation and lipid metabolism through dual regulation of adipogenesis and lipolysis in 3T3-L1 adipocytes

doi: 10.1080/19768354.2026.2623321

Figure Lengend Snippet: Schematic representation of the proposed mechanism by which Quercus infectoria gall (QIG) extract modulates adipocyte hyperplasia and hypertrophy. The extract inhibits adipocyte differentiation by inducing G 1 /S cell cycle arrest and suppressing mitotic clonal expansion in preadipocytes. It also downregulates adipogenic regulators (C/EBPβ, C/EBPα, PPARγ) and effectors (FAS, FABP4), leading to reduced differentiation and lipid accumulation. In mature adipocytes, QIG extract enhances lipolysis by upregulating adipose triglyceride lipase (ATGL) and promoting phosphorylation of hormone-sensitive lipase (HSL), thereby increasing glycerol and free fatty acid (FFA) release and reducing lipid droplet size. Together, these dual effects attenuate both adipocyte hyperplasia and hypertrophy, highlighting the anti-obesity potential of QIG extract.

Article Snippet: Primary antibodies used in this study included C/EBPβ (LAP) Antibody (#3087), C/EBPα (D56F10) XP® Rabbit mAb (#8178), PPARγ (81B8) Rabbit mAb (#2443), Fatty Acid Synthase (FAS) (C20G5) Rabbit mAb (#3180), and Phospho-HSL (Ser660) Rabbit mAb (#45804), all purchased from Cell Signaling Technology (Danvers, MA, USA); HSL Rabbit pAb (A15686) and ATGL/PNPLA2 Rabbit pAb (A6245) from Abclonal (Wuhan, China); and β-actin Rabbit mAb (MABT523) from Merck (Darmstadt, Germany).

Techniques: Phospho-proteomics