Alcoholic extract of dried AGN root was prepared by Kim’s group with modification at the Oriental Medical Hospital of Kyunghee University (Seoul, Korea). Instead of manual extraction (5 (link), 13 ), bulk extraction of powdered AGN root was carried out in a hospital herbal pharmacy extractor with 95% hot alcohol. Chemical fingerprinting of AGN by HPLC-UV showed that content of D plus DA in AGN extract was 60%. Since D and DA possess identical anti-AR, anti-proliferative and apoptotic cellular activities (5 (link), 6 (link), 18 (link)), and their majority is converted to DOH (8 (link)–11 (link)), they were isolated together as a mixture by Xing’s group at University of Minnesota. The purity of D/DA was > 99% based on monitoring with NMR and HPLC.
Mayer’s hematoxylin was obtained from Sigma-Aldrich (St. Louis, MO) and eosin Y was purchased from Thermo Fisher Scientific (Pittsburgh, PA). SV40-T antigen (T-Ag), androgen receptor (AR) and synaptophysin antibodies were purchased from BD Pharmingen (San Diego, CA), EMD Millipore (Billerica, MA) and BD Transduction Laboratories (San Diego, CA), respectively. PCR primers were purchased from Integrated DNA Technologies, Inc. (Coralville, IA).
Mayer’s hematoxylin was obtained from Sigma-Aldrich (St. Louis, MO) and eosin Y was purchased from Thermo Fisher Scientific (Pittsburgh, PA). SV40-T antigen (T-Ag), androgen receptor (AR) and synaptophysin antibodies were purchased from BD Pharmingen (San Diego, CA), EMD Millipore (Billerica, MA) and BD Transduction Laboratories (San Diego, CA), respectively. PCR primers were purchased from Integrated DNA Technologies, Inc. (Coralville, IA).
