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. 2010 Oct;177(4):1710–1724. doi: 10.2353/ajpath.2010.090903

Figure 9.

Figure 9

NoC1 does not increase active TGF-β or Smad2 phosphorylation. A: Levels of active TGF-β in the conditioned media of MEFs treated with 10 nmol/L of NoC1 or 10 nmol/L TSP1 in the presence of 2 μmol/L ascorbic acid for 48 hours were measured by PAI-1 luciferase assay (black bars). Collagen I ECM levels were calculated by densitometry of immunoblots for collagen I in the 4% DOC insoluble fraction of treated cells (gray bars). Results are reported as the fold change normalized to the mean ± SD. A representative collagen blot from this experiment is shown. B: Immunoblot for phosphorylated Smad 2 and total Smad 2/3 in MEF cell lysates from cells treated with media containing 2 μmol/L ascorbic acid and PBS, 10 nmol/L NoC1, or 8 pmol/L active TGF-β1 for 0, 1, 4, 8, and 24 hours. Cell lysates were harvested, separated by SDS-PAGE, and then immunoblotted for phosphorylated Smad 2. Blots were stripped and reprobed for total Smad 2/3.