length standard  (R&D Systems)

Journal: The Journal of endocrinology

Article Title: Intrinsic biological activity of the thrombospondin structural homology repeat in connective tissue growth factor.

doi: 10.1677/joe.1.06719

Figure Lengend Snippet: Figure 3 Stimulation of p44/p42 MAPK phosphorylation by CCN23. (A) 5·0105 HSC/well were cultured in 6-well plates until they were 80–90% confluent, after which the medium was replaced with serum-free DMEM for 48 h. The cells were then treated with TGF-1, CCN2, or CCN23 at the indicated doses for 30 min. Cells were harvested, lysed and subjected to Western blot with anti-p42/p44 or anti-phospho-p42/p44 antibodies. (B) Time course of p42/p44 MAPK phosphorylation following stimulation of serum-starved HSC with 50 ng/ml CCN23. (C) Serum-starved HSC were pretreated with 0–10 M PP2 for one hour and then stimulated with 50 ng/ml CCN23 for 30 min prior to Western blot detection of activated or total p42/p44 MAPK. The results are representative of three independent experiments.

Article Snippet: Controls included 50 ng/ml full-length human recombinant CCN2 (Ball et al. 2003b) or 2 ng/ml transforming growth factor beta 1(TGF- 1; R&D Systems, Minneapolis, MN, USA).

Techniques: Phospho-proteomics, Cell Culture, Western Blot

Journal: The Journal of endocrinology

Article Title: Intrinsic biological activity of the thrombospondin structural homology repeat in connective tissue growth factor.

doi: 10.1677/joe.1.06719

Figure Lengend Snippet: Figure 4 Promotion of pro-fibrogenic pathways in HSC by CCN23. 2·510 5 HSC/well were cultured in 6-well plates until they were 80–90% confluent, after which the medium was replaced with serum-free DMEM for 48 h. The cells were treated with TGF-1 (2 ng/ml), full-length CCN2 (50 ng/ml) or CCN23 (50, 100 ng/ml) in serum-free medium for 48 h at the indicated concentrations and then extracted for protein or RNA analysis by RT-PCR or Western blot, respectively. (A) FN mRNA (upper panel), -actin mRNA (middle panel) or FN protein (lower panel) and (B) Pro-collagen type IV(5) mRNA (upper panel) and -actin mRNA (lower panel).

Article Snippet: Controls included 50 ng/ml full-length human recombinant CCN2 (Ball et al. 2003b) or 2 ng/ml transforming growth factor beta 1(TGF- 1; R&D Systems, Minneapolis, MN, USA).

Techniques: Cell Culture, Reverse Transcription Polymerase Chain Reaction, Western Blot