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. 2019 Sep 30;14(9):e0223252. doi: 10.1371/journal.pone.0223252

Fig 2. DKK1 induces opposite effects on the Wnt/beta-catenin pathway and on its target genes as a result of mutations in the CTNNB1 gene.

Fig 2

(a) mRNA expression of oncogenes c-Myc, cyclin D1, MDM2, and hTERT were decreased in PLC/PRF/5 cells but increased in HepG2/C3A cells after DKK1 treatment. (b) P53, P21 and RB transcription decreased after DKK1 treatment of HepG2/C3A cells but were increased in PLC/PRF/5 cells. mRNA expression was measured with qPCR, and the expression was normalized to that of GAPDH and expressed as the mean ± SD of triplicates. Western blot (c) and densitometric analysis to calculate the normalized ratio of non-phosphorylated β-catenin (active) to β-actin (d) protein expression. Data are expressed as the mean ± SD from 3 different experiments. * p <0.05, ** p <0.01, *** p <0.001, as indicated. (e) Sanger sequencing chromatograms of exon 3 of the CTNNB1 gene shows point mutations in the HepG2/C3A cell line (g.311_312insC; g.316delA) while in PLC/PRF/5, no mutations were detected. The chromatograms were generated using ChromasPro software.