Journal: bioRxiv
Article Title: Reciprocal regulation between the protein arginine deiminases and mSWI/SNF chromatin remodelers controls skeletal muscle differentiation and regeneration
doi: 10.64898/2026.01.20.700682
Figure Lengend Snippet: (A) Representative light micrographs of primary myoblasts differentiated for 24 h and immunostained for MHC under control (untreated), DMSO, AFM32A, or GSK484 treatment. Scale bar, 280 μm. (B,C) Representative immunoblots (left) and corresponding quantification (right) of myogenin, MHC, PAD2, PAD3, PAD4, and the histone H3 citrullination mark, H3CitR2+R8+R17, in primary myoblasts differentiated for 24 h in the presence of AFM32A (640 µM; B ) or GSK484 (40 µM; C ). (D) Schematic representation of the in vivo experimental design to assess the effects of PAD inhibition on skeletal muscle regeneration. Cardiotoxin (CTX) was injected into the tibialis anterior (TA) muscle, followed by intraperitoneal administration of AFM32A or GSK484 at 3-, 4-, and 5-days post-injury (dpi), with analysis at 7 dpi in male and female mice. ( E ) Hematoxylin & Eosin (H&E) staining of representative 10 μm cross-sections of contralateral undamaged TA muscle and injured TA at 7 dpi under vehicle control, AFM32A, or GSK484 treatment. (F) Myofiber regeneration was quantified by measuring fiber diameter. Scale bars, 270 μm. (G) Representative PicroSirius Red and Masson’s trichrome staining of 10 μm TA muscle cross-sections from contralateral undamaged and injured muscles under vehicle, AFM32A, or GSK484 treatment. Bar graphs represent mean ± SEM from three independent experiments (N = 3). *p < 0.05, **p < 0.01, ***p < 0.001, and ****p < 0.0001.
Article Snippet: Primary myoblasts were cultured under differentiation conditions with the pan-PAD enzyme inhibitor BB-Cl amidine (HY-111347; MedChemExpress), the PAD2 inhibitor, AFM32A synthesized as previously described , and the PAD4 inhibitor GSK484 , (SML1658; MilliporeSigma).
Techniques: Control, Western Blot, In Vivo, Inhibition, Injection, Staining, Muscles
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