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. 2015 Oct 28;10(10):e0141589. doi: 10.1371/journal.pone.0141589

Fig 3. Hormone deprivation increased β-catenin interaction with TCF4 and β-catenin binding to TCF binding sites.

Fig 3

A, Expression of Wnt/-catenin target genes (eGFP reporter and Axin2) and the AR target gene (PSA) was analyzed. LNCaP cells were hormone-deprived for 3 days and then treated with vehicle, 10 nM DHT, 9 μM GSK-3 inhibitor or a combination of both for 24 h. B and C, -catenin or AR binding on target genes was analyzed using chromatin-immunoprecipitation. LNCaP cells were hormone-deprived for 3 days and then treated with vehicle, 100 nM DHT, 9 μM GSK3-i or a combination of both for 4 h. D and E, -catenin interaction with TCF4 or AR was analyzed using co-immunoprecipitation. LNCaP cells were hormone-deprived for 3 days and then treated with vehicle, 100 nM DHT, 9 μM GSK-3 inhibitor or a combination of both for 4 h. The protein lysates were either immunoblotted with indicated antibodies (D) or subjected to co-IP studies (E) Quantification of β-catenin and active β-catenin protein levels are shown below panels in (D). Relative densitometry is normalized to vehicle alone, set to 1. The data presented is representative of three independent experiments and the indicated error is the standard deviation.