Fig. 1.

HTS identifies DOS-derived small-molecule autophagy modulators. (A and B) HeLa cells stably expressing GFP-LC3 were treated with compounds in 8-point dose (4 h), and the number of GFP punctae per cell was quantified by fluorescence microscopy and automated image analysis. Representative images of cells treated with DMSO, PI-103 (2.5 µM), BRD5631 (10 µM), or BRD34009 (10 µM) are shown. Data points of dose curves are presented as the mean ± SD, n = 3 from a representative experiment. (C) Results of GFP-LC3 punctae formation assay show that BRD5631 analogs corroborate structure-activity relationship trends observed in the primary HTS. Data are the mean ± SD of three independent experiments. (D) Chemical structures of five prioritized compounds.