Additional file 7.
Screening approach for novel regulators of autophagic activity. Green fluorescent protein (GFP)-microtubule-associated protein 1 light chain 3 B (LC3) cells were plated in 96-well format and incubated in full medium (FM) with small compounds (10 μg/ml) for 6 h. Experiments were carried out in duplicates and cells were analyzed by flow cytometry for fluorescence intensities of GFP-LC3. Selected hits were then run through secondary analysis by multiparametric analysis of lysosomal degradation pathways, quantification of mCherry-GFP (tandem)-LC3, GFP-Rab7 (including normalization to control (Ctr) constructs) and LysoTracker Red (LTR) at various concentrations. Hits were validated by biochemical/imaging analysis.
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Hundeshagen et al. BMC Biology 2011 9:38 doi:10.1186/1741-7007-9-38