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

Journal: Arthritis Research & Therapy

Article Title: Effect of Porphyromonas gingivalis infection on gut dysbiosis and resultant arthritis exacerbation in mouse model

doi: 10.1186/s13075-020-02348-z

Figure Lengend Snippet: Effect of PgPAD on the onset of joint arthritis. The effect of PgPAD on the onset of arthritis was determined. In this experiment, PgPAD deletion mutant 33277 strain (⊿ pad ) and Pg 33277 wild-type (WT) strain were used, unless Pg W83 strain was specifically described in caption of Figures. ABL of the upper jaw in each group was measured ( a ). Ctrl, PBS inoculation; WT, Pg wild-type inoculation; ⊿ pad , Pg pad knockout strain inoculation. To evaluate inflammation of the gingival tissue 6 weeks after Pg inoculation, IL-6 production in gingival tissue of SKG mice was measured by ELISA ( b ). AS was measured 3 and 6 weeks after Pg inoculation with LA i.p. injection ( c ). To assess CP synthesis, ELISA was performed ( d gingival tissue, e joint tissue, f small intestine, g large intestine). The titer of serum ACPA 6 weeks after Pg inoculation with LA i.p. injection in mice from each group was measured by ELISA ( h ). The production of endogenous PADI2 and PADI4 in joint tissue was determined by Western blotting, and then the density of each band was measured by NIH Image-J ( i and j ). Data represent mean ± SD ( b – j ) of 5 mice per group. Statistical analyses were performed using the Tukey-Kramer test and Bonferroni-corrected Mann-Whitney U test for multiple comparisons (* P < 0.05, ** P < 0.01)

Article Snippet: As a control, the amount of Glyceraldehyde 3-phosphate dehydrogenase (GAPDH) was detected by anti-GAPDH antibody (10 μg/ml, HRP-60004, PROTEINTECH JAPAN, Japan) The density of target PADI2 and PADI4 bands was measured by NIH Image-J software.

Techniques: Mutagenesis, Knock-Out, Enzyme-linked Immunosorbent Assay, Injection, Western Blot, MANN-WHITNEY

Journal: Journal of immunology (Baltimore, Md. : 1950)

Article Title: Citrullination alters immunomodulatory function of LL-37 essential for prevention of endotoxin-induced sepsis

doi: 10.4049/jimmunol.1303062

Figure Lengend Snippet: Human MDMs (A) and mouse RAW 264.7 (B) macrophages were stimulated with 10 ng/mL LPS in the presence of native or citrullinated LL-37 at the indicated concentrations (0.1–10 μg/mL). Citrullinated LL-37 was obtained by treatment of the native peptide with human PAD2 or PAD4 at 23.3 U/mg peptide). The level of TNF-α (A) and NO (B) in the culture supernatants was determined using ELISA or the Griess assay at 6 or 20 h post-stimulation, respectively. Since neither the LL-37 nor the PAD enzymes alone induced the release of NO or TNF-α, for the sake of clarity these controls are not shown in the figure. Data represent the mean ± SD of three independent experiments. ns, not significant; *, p<0.05; **, p<0.01; ***, p<0.001.

Article Snippet: Recombinant human PAD2 and PAD4 were obtained from Modiquest (The Netherlands).

Techniques: Enzyme-linked Immunosorbent Assay, Griess Assay