Fig. 2

EGR1, as well as EGR1 response genes, were regulated by iguratimod in human dermal fibroblasts. Dermal fibroblasts from healthy donors were stimulated by TGFβ and treated with 0.1% DMSO or iguratimod in different concentrations. A Iguratimod suppressed the expression of the EGR family, particularly EGR1. Each cell represents normalized gene expression of an independent qPCR experiment. IGU, iguratimod B mRNA level of EGR1 in human dermal fibroblasts treated with TGFβ and different concentrations of iguratimod N = 3 per group. *, 0.05 > P > 0.01; **, 0.01 > P > 0.001, one-way ANOVA with Bonferroni multiple comparisons test was used for statistical analysis. Data are represented as mean ± standard deviation. C immunofluorescence staining of EGR1 together with DAPI in human dermal fibroblasts treated with TGFβ or iguratimod (N = 5 per group). Representative image (400 ×) and quantification are included. One-way ANOVA with Bonferroni multiple comparisons test was used for statistical analysis, ***, P < 0.001. Data are represented as mean ± standard deviation. D Volcano plot showed the differentially expressed genes in TGF-β stimulated fibroblast, with or without iguratimod treatment. E Iguratimod largely neutralized the effects of TGF-β on EGR1 response genes. This heatmap shows 327 EGR1 response genes that could be significantly regulated by TGF-β. F List of the top upstream regulators calculated by IPA. G Component genes of the TGF-β pathway that changed with iguratimod treatment. Red represents up-regulation and green represents downregulation