irisin concentrations  (BioVendor Instruments)

Journal: Cell Communication and Signaling : CCS

Article Title: Down-regulation of EHMT2 through irisin-mediated epigenetic modification promotes osteogenesis via promoting DLX3 transcription

doi: 10.1186/s12964-025-02463-x

Figure Lengend Snippet: Irisin down-regulates PIAS4 to repress EHMT2 SUMOylation, thereby promoting osteogenic differentiation. MSCs were infected with lentivirus containing oe-NC or oe-PIAS4 with or without irisin together. A Osteogenic differentiation potential was evaluated by ALP and ARS staining, and quantitative analysis was provided (scale bar = 100 μm). B Western blotting analysis of protein levels of PIAS4, EHMT2, DLX3, Runx2, OCN, OPN, and Colla1. (C) Cells were lysed and immunoprecipitated with EHMT2 antibody and then immunoblotted with SUMO2/3 or EHMT2 antibody. MSCs were treated with irisin together with or without irisin inhibitor RGDS. (D) Osteogenic differentiation potential was determined by ALP and ARS staining, and quantitative analysis was provided (scale bar = 100 μm). (E) The protein levels of FAK, p-FAK, SKP2, PIAS4, EHMT2, DLX3, Runx2, OCN, OPN, and Colla1 were assessed by Western blotting. (F) Cell lysates were immunoprecipitated with EHMT2 antibody, followed by immunoblotting with SUMO2/3 or EHMT2 antibody. (G) Ubiquitin level of EHMT2 was analyzed by Co-IP. n = 3. One-way ANOVA (for B, E) was performed for statistical analysis. * p < 0.05, ** p < 0.01, *** p < 0.001

Article Snippet: MSCs were treated with 100 ng/mL irisin (MCE, NJ, USA) together with or without irisin inhibitor RGDS (100 nM, MCE) for 48 h.

Techniques: Infection, Staining, Western Blot, Immunoprecipitation, Ubiquitin Proteomics, Co-Immunoprecipitation Assay