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. 2021 Sep 25;31(11):1559–1567. doi: 10.4014/jmb.2109.09002

Fig. 1. Identification of Morus alba L. root bark extract (MRBE) as an inhibitor of the Wnt/β-catenin pathway.

Fig. 1

(A) Firefly luciferase (FL) activity determined in HEK293-FL reporter cells incubated with either DMSO (vehicle) or MRBE (20 and 40 μg/ml) in the presence of Wnt3a-CM for 15 h. (B) Downregulation of β-catenin protein levels by MRBE in the presence of Wnt3a-CM. (C) Effect of MRBE on β-catenin mRNA levels. Real-time PCRs for the analysis of β-catenin and GAPDH were performed with total RNA prepared from HEK293-FL reporter cells treated with either DMSO or the indicated concentrations of MRBE, in the presence of Wnt3a-CM, for 15 h. (D) Western blot with the anti-β-catenin antibody on cytosolic proteins prepared from HEK293-FL reporter cells exposed to MRBE (20 μg/ml) for 15 h and MG-132 (10 μM) for 8 h. (E) Luciferase assay of HEK293 reporter cells incubated with MRBE in the presence of 20 mM LiCl for 15 h. (F) Western blot with the β-catenin antibody on cytosolic proteins from HEK293 reporter cells treated with DMSO or increasing amounts of MRBE in the presence of 40 mM LiCl for 15 h. The results represent the mean ± SD of three independent experiments. *p < 0.05 and **p < 0.01, cells treated with either Wnt3a-CM or LiCl compared with cells treated with MRBE.