Fig. 1

iECs derived from the C1-2 hiPSC line were analyzed for sprouting abilities. a Schematic is outlining the experimental workflow. b Representative confocal images of HUVEC (left) and iEC (right) sprouting spheroids after 24 h in collagen gel in media supplemented with or without VEGF. Arrows indicate disconnected cells. c Quantification of the average number (#) of sprouts per spheroid and d average sprout length (N = 3, n = 25–30). e Extracellular Acidification Rate measured via Seahorse Assay in C1-2 iECs and HUVECs (red, non-glycolytic acidification; green, glycolysis; purple, glycolytic max; N = 3, n = 9). f Western blot analysis of glycolysis enzymes and g quantification, N = 3. Statistical significance levels are set at *p ≤ 0.05, **p ≤ 0.01, ***p ≤ 0.001, and ****p ≤ 0.0001 by two-tailed Student’s t-test and Sidak’s multiple comparison test. For bar graphs: data are presented as mean ± SD. For box and wisker plots: centerline, median; box limits, upper and lower quartiles; wiskers, minimum to maximum value. Scale bar: 100 μm.