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Tonsil-derived mesenchymal stem cells alleviate skin inflammation by modulating neutrophil extracellular trap formation and T cell migration

Fig 9

NET formation analysis.

(A) Representative images of differentiated HL-60 (dHL-60) cells used as neutrophil-like cells for in vitro experiments. (B) Quantification of NETosis in dHL-60 cells under various treatment conditions: untreated, DNCB-treated, co-cultured with T-MSCs, or co-cultured with T-MSCs and DNCB. Data were analyzed using two-way ANOVA, followed by Tukey’s post hoc test for multiple comparison (*P < 0.05 and **P < 0.01). (C) Immunoblot analysis of citrullinated histone H3 (CitH3) and β-actin in dHL-60 cells treated with PMA, T-MSCs, PAD4 inhibitor (GSK484) and NADPH oxidase inhibitor (DPI) for NETosis quantification. (D) Densitometric quantification of CitH3 expression normalized to β-actin. Bars represent mean ± SD of three independent experiments. Data were analyzed using two-way ANOVA, followed by Tukey’s post hoc test for multiple comparison (* P < 0.05). ns: not significant.

Fig 9

doi: https://doi.org/10.1371/journal.pone.0343617.g009