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Image Search Results
Journal: Oncogene
Article Title: Extracellular lumican augments cytotoxicity of chemotherapy in pancreatic ductal adenocarcinoma cells via autophagy inhibition.
doi: 10.1038/onc.2016.20
Figure Lengend Snippet: Figure 7. Extracellular lumican increases gemcitabine-induced mitochondrial damage, leading to ROS accumulation, depolarization of mitochondrial membrane potential and Cyto C release. (a) MDA-PATC53 and PANC-1 cells were treated with 1 μM gemcitabine in the absence or presence of 2 μg/ml lumican for 24 h and then were analyzed by western blot. (b) Tumor tissue lysates from wild-type and lumican knockout mice were subjected to western blot. (c) The indicated cells were treated as described in a, and ROS production was identified in live cells by using flow cytometry. (d) MDA-PATC53 cells were treated with 1 μM gemcitabine or/and 2 μg/ml lumican for 24 h. Mitochondria were isolated from the cells using the Mitochondria Isolation Kit (Thermo Scientific, Rockford, lL, USA) and subjected to western blot. The bar graphs in the upper panel are the densitometric values of cytochrome c (Cyto C) in each lane. (e) MDA-PATC53 cells were treated as described in d, and isolated cell cytosol lysates were subjected to immunoblot analyses (left) and immunoprecipitated by Cyto C, followed by immunoblot analysis (right). (f) In MDA-PATC53 cells treated as described in d, depolarization of mitochondrial membrane potential in live cells was shown with Tetramethylrhodamine, methyl ester (TMRM) staining. (g) This schematic model depicts that lumican reduces chemotherapy-induced autophagy through inhibition of AMPK, augmenting cytotoxicity of chemotherapy in PDAC cells.
Article Snippet: Mitochondrial membrane potential assay Depolarization of mitochondrial membrane potential in live cells was measured by Tetramethylrhodamine,
Techniques: Membrane, Western Blot, Knock-Out, Cytometry, Isolation, Immunoprecipitation, Staining, Inhibition