Function of DEPTOR in angiogenic responses in vitro. (A) Spheroids derived from either control or DEPTOR siRNA-transfected HUVECs were embedded in a collagen I matrix, cultured for 24 hours, and stained with Alexa Fluor 488–conjugated phalloidin. A representative image of each group is shown without (upper panels) and with (lower panels) staining. The bar graph shows quantitative analysis of the mean total sprout length (±SEM) performed on ≥5 spheroids per experimental group in 3 independent experiments (**P < .01). (B) HUVECs were transfected with control or 2 DEPTOR siRNAs and cultured for 48 hours until confluent. Subsequently, linear scratch/wounds were created in the monolayers with a pipet tip, and the migration of cells into the wound was measured after 16 hours in the absence or presence of rapamycin (10 ng/mL) and/or U0126 (10 μM). Illustrated are representative photomicrographs of wounds at 0 hours and after 16 hours; dotted lines highlight the linear scratch/wound for each group of cells (representative of 3 experiments). The bar graph shows the mean percentage wound closure in pooled DEPTOR siRNA-transfected cells vs controls (±SEM; *P < .05 vs untreated control siRNA-transfected ECs; ^#P < .05 vs untreated DEPTOR siRNA-transfected ECs).