Fig. 2

AR was secreted into the extracellular medium upon VPA treatment. a Cells were treated with or without 500 μM VPA for 48 h. The serum-free conditioned media were harvested as described in the Materials and Methods, and analyzed by a human growth factor antibody array. Left panel: Template shows the location of specific antibodies spotted on the membrane. Each antibody was spotted in duplicate. Right panel: The relative density of the spots was quantified by densitometry analysis. The histograms illustrate the relative amount of secretion proteins. b Secreted levels of AR, EGFR, TGF-2β, and VEGF were analyzed in cell culture media using Western blot. Equal protein loading was confirmed by membrane staining with Ponceau S. The results are representative of at least 3 independent experiments. c-d Cells were treated with the indicated concentrations of VPA (0–500 μM) for 48 h. The mRNA levels of AR in cells were measured by real-time PCR. Intracellular AR levels were analyzed in whole cell lysates using Western blot. α-Tubulin was used as a loading control