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. 2016 Aug 4;7(35):56699–56712. doi: 10.18632/oncotarget.11057

Figure 2. Constitutively active nuclear YAPdc restricts cellular quiescence in 5F31.

Figure 2

(A, B) Ectopic expression of YAPdc by RT-qPCR and Western blot in control vector (Ctrl)- and YAPdc-transfected 5F31 cells. (C) Immunoblots using antibodies directed against YAP, the Flag epitope and Sp1 in nuclear (N) and cytoplasmic (C) fractions from untreated and 5FU-treated (40 μM, 96 hours) YAPdc and 5F31 cells. (D) transcriptional activity of the TEAD-luciferase reporter gene in control and YAPdc 5F31 cells. (E and F) Expression of the YAP target genes Cyr61, CTGF, AXL and ANKRD1 by RT-qPCR and Western blotting in control and YAPdc 5F31 cells. (G) Percentage of cells in the G0 phase after 96 hour 5FU-treatment (40 μM) in control and YAPdc 5F31 cells. Control vector and YAPdc-transfected cells were incubated for 96 hours in the presence or absence of 5FU and processed by flow cytometry analysis for quantification of G0 phase cells using Ki-67 labelling. Data are from 5 replicates. (H) Colony forming units assay of untreated and 5FU-treated YAPdc and control 5F31 cells. Western blots are representative of 3 experiments.