Fig. 2

HTS reveals luteolin as a single hit to increase mitochondrial bioenergetic function. a Pipeline of the HTS design and validation for compounds targeting mitochondria. b 10 P-differentiated SH-SY5Y cells were incubated with 1200 Prestwick library-selected compounds (10 μM, 24 h), which were automatically transferred to 384-well plates using an acoustic dispenser. Total ATP levels were measured, and hits were selected when ATP z-scores ≥ 2 and cytotoxicity z-scores ≤ 2 (green dots). Luteolin is marked in red. c Heatmap representing the ATP z-scores of selected hits from B in a 3-CRC (24 h incubation). Arrow heads represent compounds with highest z-scores (> 6) or dose-dependent increase in ATP z-scores that were selected to be tested in a 9-CRC in d. d Quantification of ATP levels and cytotoxicity in a 9-CRC (24 h incubation) of the selected hits from c. Green background indicates range of concentrations with observed increases in ATP levels, while red background indicates increased cytotoxicity