hek293t pad2 cells  (ATCC)

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) Confocal micrographs of primary myoblasts differentiated for 24 h and immunostained with anti-PAD2 (red) and anti-PAD3 or anti-PAD4 (green) antibodies. Nuclei were counterstained with DAPI (blue). Scale bar, 5 μm. (B) Subcellular fractionation of primary myoblasts differentiated for 24 h showing whole-cell (WC), cytosolic (Cyt), and nuclear (Nuc.) fractions. Representative immunoblots (left) and corresponding quantification (right) of PAD2, PAD3, and PAD4 protein levels are shown. Lamin A/C and α-tubulin were used as purity controls for the nuclear and cytoplasmic fractions, respectively. 20 ug of each extract was loaded onto the gels. (C) Representative immunoblots (left) and quantification (right) of histone H3 citrullination marks (H3CitR2+R8+R17, H3CitR17, and H3CitR26) in proliferating (P or Prol), time 0 (0 h), or differentiating (24 h) primary myoblasts. (D-G) ChIP-qPCR analysis showing enrichment of H3CitR2+R8+R17 (D) , H3CitR26 (E) , H3CitR17 (G) and total H3 (H) at the Pax7 promoter and at regulatory sequences controlling the indicated myogenic genes in proliferating (Prol), time 0 (0 h), and differentiating (24 h) primary myoblasts. The IgH enhancer was included as a negative sequence control. Bar graphs represent mean ± SEM from three independent experiments. *p < 0.05, **p < 0.01, ***p < 0.001, and ****p < 0.0001. n.s., not significant.

Article Snippet: Rabbit anti-BAF250A (A18650) was from Abclonal Technologies Rabbit anti-PAD2 (E3P8Z; #97647), anti-vinculin (E1E9V; #13901), anti-Histone H3 (#9715), Histone H3Cit R17 (E4O3F; #97272), and anti-DPF2 (E7N8J; #71642) were from Cell Signaling Technologies.

Techniques: Fractionation, Western Blot, ChIP-qPCR, Sequencing, Control

Journal: Scientific Reports

Article Title: Peptidylarginine deiminase 2 contributes to pathogenesis in trinitrobenzenesulfonic acid-induced colitis through macrophage extracellular trap-independent pathways

doi: 10.1038/s41598-025-24221-2

Figure Lengend Snippet: Generation of PAD2KO mice using the CRISPR-Cas9 systems and pathogenesis of TNBS-induced colitis in wild-type and PAD2KO mice. ( a ) The sequences produced for the generation of PAD2KO mice using the CRISPR-Cas9 systems, deficient in 61-bp on exon2 of the genomic PAD2 DNA. ( b ) PAD2 mRNA level was determined in intraperitoneal macrophages by qRT-PCR normalized to TBP. Data were presented as the means ± S.E.M. (n = 8). ****P < 0.0001 vs. wild-type (WT) mice. ( c ) PAD2 expression was determined in intraperitoneal macrophages by western blotting normalized to β-actin expression. Data are presented as the means ± S.E.M. (n = 3). ***P < 0.001 vs. WT mice. ( d ) Body weight was measured daily in untreated-, vehicle (30% ethanol)-, or TNBS-injected WT and PAD2KO mice. On day 3, colon length was measured ( e ), representative images were obtained ( f ), and macroscopic analyses of the injured area were carried out using Image J ( g ). Scale bars are 10 mm. Histological changes were observed following H&E staining ( h ). ( i ) MPO activity was determined in those mice. Data are presented as the means ± S.E.M. (n = 8–9). #### P < 0.0001 vs. vehicle-injected mice (WT EtOH), and ****P < 0.0001, ***P < 0.001, *P < 0.05 vs. TNBS-injected mice (WT TNBS).

Article Snippet: The membranes were then incubated with PAD2 antibody (1:500) (Proteintech, 12110-1-AP) and β-actin antibody (1:2000) (CST, #4970).

Techniques: CRISPR, Produced, Quantitative RT-PCR, Expressing, Western Blot, Injection, Staining, Activity Assay

Journal: Scientific Reports

Article Title: Peptidylarginine deiminase 2 contributes to pathogenesis in trinitrobenzenesulfonic acid-induced colitis through macrophage extracellular trap-independent pathways

doi: 10.1038/s41598-025-24221-2

Figure Lengend Snippet: Expression of inflammatory cytokines, chemokines, and PAD2 in PAD2KO mice. ( a ) TNFα, ( b ) IL-1β, ( c ) IL-12, ( d ) IL-23, ( e ) IFNγ, ( f ) CXCL2, and ( g ) PAD2 mRNA expression were determined using qRT-PCR in colon tissues of vehicle (30% ethanol)-, or TNBS-injected WT and PAD2KO mice on day 3, and mRNA levels were normalized to TBP. Data are presented as the means ± S.E.M. (n = 8–10). #### P < 0.0001, ### P < 0.001, ## P < 0.01 vs. vehicle-injected mice (WT EtOH), and ****P < 0.0001, *P < 0.05 vs. TNBS-injected mice (WT TNBS).

Article Snippet: The membranes were then incubated with PAD2 antibody (1:500) (Proteintech, 12110-1-AP) and β-actin antibody (1:2000) (CST, #4970).

Techniques: Expressing, Quantitative RT-PCR, Injection