Fig. 5

Time course of STO-609-mediated inhibition of autophagic flux in OVCAR8 mCherry-eGFP-LC3B spheroids. a OVCAR8 mCherry-eGFP-LC3B cells were pretreated with either 10 μM STO-609 or DMSO and seeded at a density of 2000 cells/well into 96-well round-bottom ULA plates. Images of eGFP and mCherry fluorescence signals were captured using the IncuCyte® ZOOM live-cell imaging system at 3h intervals over a period of 14 days. Representative images are shown for specific time points (days 2–12). Scale bar = 200 μm. b Quantification of eGFP, mCherry and Overlap fluorescence signals were quantified from 10 independent wells using the IncuCyte® ZOOM image analysis software. Data (mean ± SD) are displayed starting at day 2 to allow for complete aggregation of cells into individual spheroids, and to avoid the increased background fluorescence signal on image edges in the GFP channel at these early time points