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. 2020 Aug 18;15(8):e0232917. doi: 10.1371/journal.pone.0232917

Fig 3. Sunitinib inhibits TGF-β1-induced Wnt signaling.

Fig 3

(A) Effect of sunitinib on TCF4 transcriptional activity. A549 cells were treated with TGF-β1 (5 ng/mL) or its combination with sunitinib for 48 h before measuring TOPflash luciferase activity. FOPflash-normalized TOPflash luciferase activity is represented as the relative TCF/LEF luciferase activity. (B) A549 cells were pretreated with sunitinib for 3 h, and then incubated with TGF-β1 (5 ng/mL) for 1 h. After RNA extraction and cDNA synthesis, the mRNA expression of TNIK and Wnt target genes was measured by qRT-PCR. (C) The protein levels of TNIK and β-catenin in cytosolic and nuclear fractions were measured by Western blot. Actin and lamin B1 were used as loading controls for the cytosolic and nuclear fractions, respectively. The results are representative of triplicate experiments. # p < 0.01 versus control; * p < 0.05, ** p < 0.01 versus treatment with TGF-β1 only.