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. 2016 Mar;186(3):678–690. doi: 10.1016/j.ajpath.2015.11.003

Figure 7.

Figure 7

Dexamethasone (dex) and bortezomib do not affect thrombospondin1 (TSP1) stimulation of transforming growth factor (TGF)-β activity or TGF-β signaling by heparanase expressing CAG (CAG-hpse) cells. CAG-hpse cells (2 × 105) were seeded overnight in RPMI with 0.5% fetal bovine serum and then treated for 1 hour with 50 or 100 nmol/L dexamethasone or 5, 50, or 100 nmol/L bortezomib. A and B: Cells then were treated for 1 hour with either 100 pmol/L TGF-β (A) or 20 μg/mL TSP1 (B). Cell lysates were analyzed for phospho-Smad2 by Western blot analysis. Densitometric analyses are normalized to β-actin. Results are the means ± SD of five to six separate experiments. C: CAG-hpse cells (5 × 105) were treated with 100 pmol/L TGF-β, 50 or 100 nmol/L dexamethasone, or 5, 50, or 100 nmol/L bortezomib in RPMI with 1% fetal bovine serum for 6 hours. RNA was harvested and analyzed for THBS1 mRNA by RT-PCR. THBS1 levels were normalized to GAPDH. D: Cells were treated as above in serum-free media for 12 hours and cell lysates harvested for Western blot analysis for TSP1. Densitometric analysis was performed and bands were normalized to β-actin. For C and D, results are the means ± SD from three separate experiments. ∗P < 0.05 versus untreated, analysis of variance with Tukey analysis.