LC3 activation was assessed with the Cyto-ID Autophagy Detection kit (Enzo) in live cells using fluorescence microscopy, according to the manufacturer’s instructions. Briefly, cells were seeded into six-well plates at a density of 1.25 million per well and treated immediately with vehicle/Dox (2 μg/ml) for 48 hours for Tet On induction, or the next day with vehicle/5 μM SoSe for 3 hours. After this 48- or 3-hour treatment, cells were washed, stained with Green autophagosome detection reagent and Hoechst 33342 nuclear stain, and analyzed by confocal microscopy using fluorescein isothiocyanate and Hoechst filters.

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