The FBG domain of TNX activates latent TGF-β in vitro. (A) Immunoblots showing phospho-Smad2 (P-Smad2) levels in NMuMG cells incubated for 3 h on either noncoated (−) or 222 pmol/cm2 FBG-coated (+) dishes or stimulated for 3 h with 5 ng/ml of soluble human TGF-β1 in the presence of 10 µg/ml cycloheximide or the corresponding vehicle (DMSO). (B) Immunoblots showing levels of phospho-Smad2 in NMuMG cells incubated for 3 h on either noncoated (−) or 222 pmol/cm2 FBG-coated (+) dishes or stimulated for 3 h with 5 ng/ml TGF-β1 in the presence of a control IgG or 10 µg/ml anti-pan–TGF-β antibody. (C) Firefly luciferase activity in MCF10A-TRE-Luc cells cultured for 16 h under the same conditions as described in B. *, P < 0.05 versus control IgG. Error bars are means ± SD. (D) F-actin direct fluorescence (top) and indirect immunofluorescence staining of E-cadherin (bottom) performed in NMuMG cells cultured for 36 h under the same conditions as described in B. Bars, 15 µm.
