Biomolecules & Therapeutics : eISSN 2005-4483 / pISSN 1976-9148

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Fig. 1. TGase 2 regulates autophagy in cancer cells. (A) Immunoblottings of autophagy markers in cancer cells showed TGase 2 is associated with autophagy in cancer. CAKI-1 and A549 cells were transfected with siRNA of TGase 2 for 24 h while MCF7 and HCT116 cells were transfected with HA-TGase 2 plasmid for 24 h. Before harvest cells, cells were treated with chloroquine (CQ, 50 µM) for 6 h under amino acid deprivation for 4 h. Whole-cell lysates were subjected to the immunoblotting with indicated antibodies. (B) Autophagy activity was regulated by TGase 2 in cancer cells. Under the same conditions as above (A), cells were stained with CYTO-ID green detection reagent for autophagy activity analysis. Fluorescence was measured at wavelength 488 nM. (C) Confocal microscopy images showed that induction of LC3 (green) is regulated by TGase 2 expression in A549 and MCF7. (D) TGase 2 knock down reduced autophagy in A549. GFP-LC3 stable expressed A549 cells were transfected with TGase 2 siRNA for 24 h. (E) TORIN-2 induced autophagy was significantly reduced by TGase 2 siRNA treatment. GFP-LC3 puncta were analyzed by live imaging using confocal microcopy. Cells were counterstained with DAPI to visualize the nuclei. Cumulative data from three independent experiments is shown here as mean ± SD (n=3). ***p<0.001, Scale bar: 20 µm.
Biomolecules & Therapeutics 2019;27:34~40
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