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. Author manuscript; available in PMC: 2020 May 29.
Published in final edited form as: Sci Immunol. 2019 Nov 29;4(41):eaax7965. doi: 10.1126/sciimmunol.aax7965

Fig. 5. Impaired response to TGF-β in the patients’ fibroblasts.

Fig. 5.

(A) Immunofluorescence of fibronectin (FN), type V collagen (COLLV), type III collagen (COLLIII), α2β1, α5β1, and αvβ3 integrins, α-smooth muscle actin (α-SMA), and cadherin-11 (CAD-11) in primary fibroblasts from a healthy control (C), P2, a patient with hEDS (hEDS) (56), and a patient with cEDS (cEDS) (93). Scale bar: 10 μm. (B) In vitro scratch assay with primary fibroblasts from a healthy control (C), P2, a patient with hEDS (hEDS) (56), and a patient with cEDS (cEDS) (93). Images were captured at 0 and 48 h after scratching. Scale bar: 100 μm. (C) Transwell assay with primary fibroblasts from a healthy control (C), P2, a patient with hEDS (hEDS) (56), and a patient with cEDS (cEDS) (93). (D) mRNA induction in primary fibroblasts from healthy controls (C1 and C2) and patients (P2 and P3) stimulated with TGF-β (10 ng/mL) for the indicated times. (E) The top 10 upregulated or downregulated genes in terms of absolute fold change, in primary fibroblasts from healthy controls (C1 and C2) stimulated with TGF-β (10 ng/mL) for 2, 6, and 24 h, with a greater than 1.5-fold change relative to patients (P2 and P3) at each time point. (F and G) Expression of JNK1 protein (F) and production of fibronectin (top panel) and IL-11 (bottom panel) (G) by primary fibroblasts from healthy controls (C1 and C2) transfected with control siRNA (50 nM) or MAPK8 siRNA (50 nM) for 48 h and then stimulated with TGF-β (10 ng/mL) for an additional 24 h. NS, non-stimulated conditions. The values shown are the means ± SEM of two (C) or three (D and G) independent experiments. *, P < 0.05, ***, P < 0.001, and ****, P < 0.0001; in unpaired t tests (D and G).