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Figure 4 | Cell & Bioscience

Figure 4

From: Up-regulation of the Hippo pathway effector TAZ renders lung adenocarcinoma cells harboring EGFR-T790M mutation resistant to gefitinib

Figure 4

TAZ mediates EMT of gefitinib-resistant PC9/GR. (A) TAZ knockdown in PC9/GR cells resulted in a morphology change in PC9/GR by a light microscope. The spindle form of the PC9/GR-shNC cells changed to a round shape. The cell density of the clusters was obviously enhanced in the PC9/GR-shTAZ cells. (B) Cell lysates from PC9/GR and PC9 cells expressing vector, shTAZ and PEX2-TAZ were separated and probed with antibodies for epithelial marker and mesenchymal markers as indicated. TAZ knockdown in PC9/GR cells increased E-cadherin and decreased vimentin, CTGF and AXL. All data were normalized to GAPDH. (C) Luciferase reporter assay in 16HBE cells with TAZ or TAZ-S51A overexpression. 16HBE cells co-transfected with pGL3-CTGF/pGL3-AXL and pEX2-TAZ showed significantly elevated reporter activity compared with controls. Meanwhile, cells co-transfected with pEX2-TAZ-S51A, which disrupts the TEAD binding, exhibited decreased luciferase activity compared with pEX2-TAZ. Cells were harvested 48 h after transfection for measurement of luciferase activity in all experiments. (D) Luciferase reporter assay in PC9/GR cells. Total amounts of DNA and RNA were kept constant. Results are expressed as means ± SEM from three independent experiments. Statistical analyses were carried out using Student’s t-test.

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