Fig. 2
Down regulation of β-catenin expression and inhibition of β-catenin signaling in HuH6pTER-sh-β-catenin transfectant cells. a HuH6 cells were transfected with pTER-sh-β-catenin and selected with 2 μg/ml puromycin for 4 weeks. After amplification of isolated colonies, resultant HuH6pTER-sh-β-catenin transfectants were cultivated in the presence or absence of 2 μg/ml doxycycline for 72 h and the abundance of β-catenin in whole cell extracts was assessed by immunoblotting. b Clone 5G was cultivated in the presence or absence of 2 μg/ml doxycycline for the indicated times and β-catenin mRNA levels were analyzed by RT-qPCR. c- Clone 5G was cultivated in the presence or absence of 2 μg/ml of doxycycline for 72 h and Axin2 mRNA levels were analyzed by RT-qPCR. d Clone 5G was cultivated in the presence or absence of 2 μg/ml doxycycline for 72 h and β-catenin/TCF transcriptional activity was assessed by the luciferase reporter assay