Role of Heparan Sulfate 2-O-Sulfotransferase in Prostate Cancer Cell Proliferation, Invasion, and Growth Factor Signaling
Figure 4
2OST modulates growth factor signaling in cell line model of prostate cancer progression. (a) Decreased FGF signaling in cells treated with 2OST siRNA. Western blotting for phospho-ERK and β-actin performed as described in Materials and Methods. Levels of p-ERK are normalized to β-actin. Densitometry figures are shown below each sample and are representative of two independent experiments. (b) Decreased TGFβ signaling in cells treated with 2OST siRNA. Levels of p-SMAD2 normalized to total SMAD2. (c) Decreased Wnt signaling in cells treated with 2OST siRNA. Levels of β-catenin normalized to β-actin. (d) Decreased SHH signaling in cells treated with 2OST siRNA. Real-time PCR analysis of SHH pathway response genes PTCH and GLI1 was performed. Error bars indicate standard deviation. Asterisk indicates . (e) Decreased complex formation between SHH and Pln. Pln was immunoprecipitated from conditioned media from either control siRNA or 2OST siRNA cells. Left pane shows western blot for SHH bound to equal amounts of Pln. Right panel shows equal amounts of SHH in input samples from coimmunoprecipitation.
