Figure 1.
Figure 1. Gene silencing of Fli1 leads to the induction of SSc-like protein and gene expression profiles in keratinocytes. (A) Immunohistochemistry for Fli1 on the skin sections of healthy controls (n = 8), lcSSc patients with anticentromere antibody (n = 8), and dcSSc patients with anti–topoisomerase I antibody (n = 5). Representative images are shown. Bars, 100 µm. (B) The results of Fli1 staining intensity in A were semiquantitatively evaluated with a four-point grading scale. Kruskal-Wallis test followed by Dunn’s posthoc test was used. (C) mRNA was isolated from skin epidermal sheets from healthy controls (n = 4) and dcSSc patients (n = 6), and gene expressions were analyzed by qRT-PCR. (D) Whole cell lysates from cultured NHKs treated with control nonsilencing SCR or FLI1 siRNA (siFLI1) were subjected to immunoblotting. The values below each blot represent the relative levels of target molecules normalized by controls with densitometry. Representative images of the blots of four independent experiments are shown. (Right) The bar graphs summarize the relative values of the density from these four experiments. (E) The mRNA expression of FLI1, SNAI1, and CDH1 in NHKs treated with SCR or siFLI1 was evaluated. Relative expression levels from four independent experiments were normalized to the expression levels treated with SCR. (F) ChIP assay in NHKs was performed with anti-Fli1 antibody and the primers specific for the designated area of the SNAI1 gene promoter. (G) The back skin taken from Fli1+/− mice and their littermate wild-type (Fli1+/+) mice were immunohistochemically stained for K6 and K16. n = 5 for each group. (Left) Representative images of immunohistochemistry are shown. Bars, 100 µm. (Right) The graphs show the results of semiquantitative scoring of the staining intensity. Data are shown as mean ± SEM. *, P < 0.05; **, P < 0.01; by two-tailed Mann-Whitney U test, unless otherwise indicated. The results are from representative experiments that have been repeated twice independently or are otherwise indicated. AU, arbitrary units; Ctrl, control.

Gene silencing of Fli1 leads to the induction of SSc-like protein and gene expression profiles in keratinocytes. (A) Immunohistochemistry for Fli1 on the skin sections of healthy controls (n = 8), lcSSc patients with anticentromere antibody (n = 8), and dcSSc patients with anti–topoisomerase I antibody (n = 5). Representative images are shown. Bars, 100 µm. (B) The results of Fli1 staining intensity in A were semiquantitatively evaluated with a four-point grading scale. Kruskal-Wallis test followed by Dunn’s posthoc test was used. (C) mRNA was isolated from skin epidermal sheets from healthy controls (n = 4) and dcSSc patients (n = 6), and gene expressions were analyzed by qRT-PCR. (D) Whole cell lysates from cultured NHKs treated with control nonsilencing SCR or FLI1 siRNA (siFLI1) were subjected to immunoblotting. The values below each blot represent the relative levels of target molecules normalized by controls with densitometry. Representative images of the blots of four independent experiments are shown. (Right) The bar graphs summarize the relative values of the density from these four experiments. (E) The mRNA expression of FLI1, SNAI1, and CDH1 in NHKs treated with SCR or siFLI1 was evaluated. Relative expression levels from four independent experiments were normalized to the expression levels treated with SCR. (F) ChIP assay in NHKs was performed with anti-Fli1 antibody and the primers specific for the designated area of the SNAI1 gene promoter. (G) The back skin taken from Fli1+/− mice and their littermate wild-type (Fli1+/+) mice were immunohistochemically stained for K6 and K16. n = 5 for each group. (Left) Representative images of immunohistochemistry are shown. Bars, 100 µm. (Right) The graphs show the results of semiquantitative scoring of the staining intensity. Data are shown as mean ± SEM. *, P < 0.05; **, P < 0.01; by two-tailed Mann-Whitney U test, unless otherwise indicated. The results are from representative experiments that have been repeated twice independently or are otherwise indicated. AU, arbitrary units; Ctrl, control.

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