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GFP-LC3 puncta assay

AN April Nettesheim
MS Myoung Sup Shim
JH Josh Hirt
PL Paloma B. Liton
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To monitor autophagy by live cell imaging, were transduced human TM cells with 5 pfu/cell of the replication-deficient adenovirus AdGFP-LC3 (kindly provided by Dr. Wen-Xing Ding, University of Kansas Medical Center). AdGFP was used as a control. Three days after transduction, the cell culture media was changed with TGFβ2 (10 ng/mL)-containing media and the cells were further incubated at 37 °C, 5% CO2 for 48 h. For starvation condition, the cell culture media was changed with Hank’s balanced salt solution (HBSS, GIBCO) and it was incubated for 6 h. Images were acquired with CELENA® S Digital Imaging System (Logos biosystems) and the images were processed by using Fiji, an image processing package.

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