Fig. 1

TMPRSS4 promotes anchorage-independent growth of prostate cancer cells. A PC3 cells were stably transfected with TMPRSS4 expression vector or empty vector. Cells were seeded in 6-well plates at a density of 5 × 10⁴ cells/well and incubated for 48 or 72 h. Cells were counted to measure proliferation. B Soft agar anchorage-independent growth assay. Transfected PC3 cells were seeded in triplicates in 6-well plates containing semi-solid agar at a density of 3 × 10³ cells/well and allowed to grow for 14 days. Representative images are shown. Total colonies (> 0.1 mm) and colonies with diameter > 0.3 mm were counted. Bar, 500 μm. C Transfected PC3 cells were incubated for 48 h, and lysed for immunoblot analysis. Anti-myc antibody was used to detect myc-tagged TMPRSS4. Densitometric quantification was performed on the immunoblots using GAPDH as a loading control except that phospho-AKT was normalized against total AKT. The mean relative density from three independent experiments is shown under the immunoblots. D Cell adhesion assay. Transfected PC3 cells were plated onto fibronectin-coated plates for 20 or 60 min. The number of adherent cells was counted in five representative fields (× 200). Values represent mean ± standard deviation (SD). ***P < 0.001