Fig. 1. High-throughput screening for natural compound-based autophagy modulators.
A Detection of starvation-induced and bafilomycin A1-inhibited autophagy in a flow cytometric screening. MEFs stably expressing mCitrine-LC3 were cultured either in full medium plus serum or in starvation medium and were treated with either 10 nM bafilomycin A1 or mock treatment (DMSO) for 6 h (n ≥ 19). The mCitrine fluorescence is shown as a percentage relative to that of mock-treated cells. Data represent means ± SEM. B, C High-throughput autophagy screening of 300 natural compounds. The diagram shows dual-flashlight plots for strictly standardized mean difference (SSMD) versus the average percentage of mCitrine-LC3. Dotted lines define potential inducers and inhibitors of autophagy. Cells were treated with 10 µM of each compound for 6 h (n = 3). Arzanol is highlighted in red. D The diagram shows individual levels of mCitrine-LC3 fluorescence from (B, C) for each of the 300 compounds in full or starvation medium. Dotted lines mark the ±15% difference in mCitrine-LC3 levels. Data represent means ± SEM.