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
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