Skip to main content
. 2015 Jan 31;6(8):6136–6150. doi: 10.18632/oncotarget.3346

Figure 3. HBC inhibits AR transcriptional activity.

Figure 3

A) Exponentially growing LNCaP cells were treated with increasing concentrations of HBC for 24 hours, and AR, PSA and β-actin levels in cell extracts were determined. B) Exponentially growing LNCaP cells were treated with 40 μM HBC or Casodex for 24 hours and RT-PCR performed to evaluate PSA, NKX3.1 and GAPDH mRNA expression. The relative density of GAPDH normalized PSA and NKX3.1 bands was determined using EagleSight software (version 3.2; Stratagene). C) HBC disrupts AR-CaM interaction. Exponentially growing LNCaP cells were treated with 40 μM HBC or Casodex for 24 hours, AR and CaM immunoprecipitates were prepared and Western blot analysis was performed as described in Materials and Methods. Input represents 10% of the protein used for immunoprecipitation. Observations are representative of 3 independent experiments. D) HBC inhibits AR binding to androgen response elements. EMSA reactions were carried out in the presence of 40 μM HBC or Casodex (CDX), or vehicle (control) and subjected to non-denaturing gel electrophoresis. EMSA was performed as described in Materials and Method. The data is representative of 2 independent experiments. Density of bands in each lane were determined by using the ImageJ program and relative densities were calculated after subtracting the background density of bands in DU145 lane from the density of bands in LNCaP lanes.