Fig. 3

Screening for small molecule modulators of Lgr5 surface expression. a Infrared image of a stable U2OS cell line expressing MarsCy1-Lgr5 in a 384-well plate without (–K44A) and with transient transfection (+K44A) of Dynamin I K44A (NT, parental U2OS cells). b Quantification of (a) and Z’-factor analysis. c Infrared image of a stable U2OS cell lines expressing MarsCy1-Lgr5 compared to MarsCy1-Lgr5/V2R-tail (NT, parental U2OS cells). d Quantification of (c) and Z’-factor analysis. e A total of 91 hits were cherry picked and incubated overnight at 37 °C on stable MarsCy1-Lgr5-EGFP cells. Black bar, wild-type Lgr5; Hatched bar, +K44A control; Pink bar, autofluorescent compounds (violet, yellow, blue, and green bars as in 3f). Hits were measured against the ± 1, 2, 3 standard deviations from wild-type DMSO mean (green, blue, and red lines). Each compound is described according to plate ID, library name (JH, John’s Hopkins; PT, Prestwick; KG, Kinase Gold), common drug name, and position on the secondary screening plate. f Synthetic glucocorticoid receptor agonists from (e) were purchased, in addition to dexamethasone, and screened in a dose-response assay. g and h Spiperone and glucocorticoids, respectively, increase plasma membrane expression of D2R-DRY and Lgr5