Quantifying Autophagy in PC3 Cells

PC3 cells were seeded in 24-well plates at a density of 8 × 10⁴ cells/well and transfected with a GFP-tagged LC3 plasmid (GFP-LC3; Addgene plasmid # 11546)52 (link) using FuGENE^® HD Transfection Reagent (Roche). Twenty hours after transfection, PC3 cells were treated as indicated and the formation of GFP-LC3 puncta in response to chemicals was observed for 48 hours thereafter using the Zeiss Axiovert 200 inverted fluorescence microscope equipped with the AxioCam HRc microscope color camera and the AxioVision 4.8 software (all from Carl Zeiss, Göttingen, DE).

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Modernelli A., Naponelli V., Giovanna Troglio M., Bonacini M., Ramazzina I., Bettuzzi S, & Rizzi F. (2015). EGCG antagonizes Bortezomib cytotoxicity in prostate cancer cells by an autophagic mechanism. Scientific Reports, 5, 15270.

Publication 2015

Axiovert 200 AxioCam HRc microscope AxioVision 4.8 software

Fluorescence microscope Fugene Microscope Pc3 cells Plasmid Transfection

Corresponding Organization :

Other organizations : University of Parma

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Variable analysis

independent variables

Treatment conditions

dependent variables

Formation of GFP-LC3 puncta

control variables

Cell line (PC3)
Transfection of GFP-LC3 plasmid
Time points for observation (48 hours)

controls

Positive control: Not specified
Negative control: Not specified

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