Fig. 2.

IL11 antibodies block TGFβ2-driven activation of skin fibroblasts from SSc patients and controls

(A) Dose–response curve and X203 IC₅₀ value in inhibiting MMP2 secretion by HDFs and SSc dermal fibroblasts stimulated with TGFβ2. (X203 serial 4-fold dilution from 4 µg/ml to 61 pg/ml; line at x = 2 µg/ml; total n = 2 per group.) (B) Transwell migration indices of HDFs following 24 h stimulation with TGFβ2 (5 ng/ml) and in the presence of IgG or X203 (2 μg/ml). Data: n = 4, mean with s.d. Statistics: one-way ANOVA, Tukey’s multiple testing correction. (C) Migration assay representative images; scale bar, 150 µm. (D) Representative images of collagen I, periostin and EdU stainings of HDFs, following a 24 h stimulation with IL11 (5 ng/ml), or a combination of TGFβ2 (5 ng/ml) and IgG (2 μg/ml), IL11 neutralizing antibody (X203, 2 μg/ml) or ERK inhibitor (U0126, 10 μM); scale bar, 100 µm. (E) Quantification of signal intensity/area (collagen I, periostin) and percentage of EdU-staining positive cells relative to experiment in (D). Statistics: one-way ANOVA, Tukey’s multiple testing correction. (F and G) Representative images and quantification of percentage of αSMA-staining positive cells following a 24 h stimulation of SSc fibroblasts with the indicated factors. Data: whiskers from min. to max. value, line at median. Statistics: one-way ANOVA, Tukey’s multiple testing correction. αSMA: α-smooth muscle actin; EdU: 5-ethynyl-2′-deoxyuridine; ERK: extracellular signal-regulated kinase; HDF: human dermal fibroblast.