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. 2012 Feb 9;7(2):e31213. doi: 10.1371/journal.pone.0031213

Figure 4. Flavokawain B (FKB) down-regulates mRNA and protein expression of AR and AR target genes.

Figure 4

(A).The protein expression of AR and PSA after indicated treatments for 16 hours or 5 µg/ml FKB for indicated period of times was analyzed by Western blot. α-Tubulin was detected as a loading control. A representative blot was shown from three independent experiments. (B).Western blotting analysis shows that FKB decreases both constitutive and a synthetic androgen, R1881, stimulated AR and PSA expression in LNCaP cell. LNCaP cells were cultured in charcoal stripped FBS medium. FKB and R1881 at indicated concentrations treated cells for 16 hours. α -Tubulin was detected as a loading control. A representative blot was shown from three independent experiments. (C). Proteasome inhibitor, MG132, does not affect FKB mediated down-regulation of AR protein expression. LNCaP cells were treated with MG132 for 2 hours and then added FKB for additional 16 hours. α-Tubulin was detected as a loading control. A representative blot was shown from three independent experiments. (D). LNCaP and C4-2B cells were treated with 5 µg/ml FKB for 4 and 8 hours. Real-time RT-PCR was performed to analyze mRNA expression of AR and AR target genes (PSA and TMPRSS2). Bars are mean ± SD of three independent experiments. FKB significantly decreases mRNA expression of AR target genes (Student t test, P<0.01).