Journal:

Article Title: Iloprost suppresses connective tissue growth factor production in fibroblasts and in the skin of scleroderma patients

doi:

Figure Lengend Snippet: Normal (gray bars) and scleroderma (black bars) fibroblasts were grown to confluence on six-well plates. The experimental conditions were as follows: C, control with medium only; I, treated with 1 ng/ml Iloprost; T, treated with 10 ng/ml TGF-β; T+I, treated with 10 ng/ml TGF-β plus 1 ng/ml Iloprost added simultaneously. Media were removed after 24 hours for assay of (a) CTGF by ELISA, and (b) N-terminal propeptide of type I procollagen by ELISA. Values shown represent the means of six replicates of each experiment using cells derived from six patients with recent-onset diffuse scleroderma and from six healthy controls. *P < 0.05; **P < 0.001. (c) Normal human fibroblasts were grown to confluence and treated with 10 ng/ml TGF-β, with or without 1 ng/ml Iloprost and then lysed after 24 hours. CTGF was assayed by Western blot analysis. (d) In other experiments normal human fibroblasts were grown to confluence and treated with 10 ng/ml TGF-β with or without 1 ng/ml Iloprost. After 8 hours, the cells were lysed, and RNA was extracted for Northern blot analysis.

Article Snippet: The nitrocellulose filters were stained with primary Ab for 1 hour using the following primary Ab’s: rabbit polyclonal anti-human CTGF as above, rabbit anti-human collagen type I (759103R; BIODESIGN International, Kennebunk, Maine, USA), mouse monoclonal IgG anti–COX-1 (SC 7299; Santa Cruz Biotechnology Inc., Santa Cruz, California, USA) goat polyclonal IgG anti–COX-2 (SC-1746; Santa Cruz Biotechnology Inc.).

Techniques: Enzyme-linked Immunosorbent Assay, Derivative Assay, Western Blot, Northern Blot

Journal:

Article Title: Iloprost suppresses connective tissue growth factor production in fibroblasts and in the skin of scleroderma patients

doi:

Figure Lengend Snippet: Normal human fibroblasts were grown to confluence in six-well plates and treated at baseline with TGF-β (gray bars) (10 ng/ml). After 12 hours, Iloprost (1 ng/ml) was added to the media of some of the cells (black bars). (a) Media were removed at various time points for type I collagen assay using Western blot analysis. (b) Additional assays were performed on the media to measure CTGF by ELISA.

Article Snippet: The nitrocellulose filters were stained with primary Ab for 1 hour using the following primary Ab’s: rabbit polyclonal anti-human CTGF as above, rabbit anti-human collagen type I (759103R; BIODESIGN International, Kennebunk, Maine, USA), mouse monoclonal IgG anti–COX-1 (SC 7299; Santa Cruz Biotechnology Inc., Santa Cruz, California, USA) goat polyclonal IgG anti–COX-2 (SC-1746; Santa Cruz Biotechnology Inc.).

Techniques: Collagen Assay, Western Blot, Enzyme-linked Immunosorbent Assay

Journal:

Article Title: Iloprost suppresses connective tissue growth factor production in fibroblasts and in the skin of scleroderma patients

doi:

Figure Lengend Snippet: Normal human fibroblasts were grown to 50% confluence and infected with an adenovirus-encoding the cDNA for CTGF at 500 viral particles per cell. After 2 hours, the media were washed off and replaced with DMEM. (a) Subsequently, media were removed for assay of CTGF (gray bars) and type I collagen(black bars), by Western blot analysis. (b) The recombinant adenovirus encoding cDNA for human CTGF. LITR, left inverted terminal repeat; RITR, right inverted terminal repeat.

Article Snippet: The nitrocellulose filters were stained with primary Ab for 1 hour using the following primary Ab’s: rabbit polyclonal anti-human CTGF as above, rabbit anti-human collagen type I (759103R; BIODESIGN International, Kennebunk, Maine, USA), mouse monoclonal IgG anti–COX-1 (SC 7299; Santa Cruz Biotechnology Inc., Santa Cruz, California, USA) goat polyclonal IgG anti–COX-2 (SC-1746; Santa Cruz Biotechnology Inc.).

Techniques: Infection, Western Blot, Recombinant