FIG. 3.

Consequences of Dio2 loss in C2C12 cells on endothelial cell function. CM was collected from WT or D2KO C2C12 differentiated for four days and used to treat HUVECs. (A) Cell number after 48 hours of growth in EBM plus the indicated additions, (B–F) branch (indicated by arrow) and mesh (indicated by a star) formation were quantitated. (G, H) Confluent HUVECs were scratched with a pipet tip, and the area remaining after regrowth ± supplementation with either WT- or D2KO-CM (indicated between black lines) was determined after 11 hours. (I, J) HUVEC migration through a transwell insert toward a lower chamber containing WT-or D2KO-CM was determined. Cells were stained with NucBlue live cell stain. Values shown are mean ± SEM. *p < 0.05, **p < 0.01, ***p < 0.001 by one-way ANOVA followed by Tukey's multiple-comparison test (A, J) or unpaired Student's t-test (C–F, H). Dio2, type 2 deiodinase; CM, conditioned medium; HUVEC, human umbilical vein endothelial cell; EBM, endothelial basal medium.