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Figure 1 | Arthritis Research & Therapy

Figure 1

From: CD109, a TGF-β co-receptor, attenuates extracellular matrix production in scleroderma skin fibroblasts

Figure 1

CD109 protein expression is increased in systemic sclerosis skin and in cultured systemic sclerosis skin fibroblasts. (A, B) Immunohistochemistry: systemic sclerosis (SSc) (Patients S1 and S19 in Table 1) and normal (controls N1 and N9) skin sections were analyzed by immunohistochemistry using an anti-CD109 antibody followed by detection with (A) Alexa Fluor 488-conjugated secondary antibody (Patient S1 and control N1) or (B) horseradish peroxidase-conjugated secondary antibody (Patient S19 and control N9) followed by diaminobenzidine staining, as described in Materials and methods. (C) Immunocytochemistry: SSc (Patient S1) and normal (control N1) skin fibroblasts were grown in culture and were analyzed by immunocytochemistry using an anti-CD109 antibody followed by detection with an Alexa Fluor 488-conjugated secondary antibody, as described in Materials and methods. Immunofluorescence microscopy was done using Olympus B202 (Carson Group Inc., Markham, ON, Canada). Images captured with a digital camera (DC-330; Dage-MTI Inc., Michigan City, IN, USA). Scale bar: 50 μM. Results presented are representative of three independent experiments. (D, E) Western blot: cell lysates prepared from cultured SSc and normal keratinocytes were analyzed by western blot using an anti-CD109 antibody. The membrane was reprobed with an anti-β-actin antibody to confirm equal protein loading (D). Densitometric analysis of CD109 protein levels (180 kDa band) in the cell lysates of SSc (n = 6) and normal (n = 6) keratinocytes was performed using β-actin as a loading control (P < 0.05).

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