Figure 7

17-AAG, EC154, and LBH589 suppress CCRCC mediated tubule formation and differentially modulate endothelial barrier function. a) Serum starved (0.1%) HUVEC cells were plated on growth factor reduced Matrigel in the presence of CCRCC conditioned medium (UMRC2 or 786-0) in the presence or absence of 17-AAG, EC154, or LBH589. Tubule formation was imaged at 6 h from 6 replicate wells of a 96-well plate (1 field per well at 40 × magnification), branch points counted, and presented as a percent of untreated control with standard deviation. Conditioned medium from both cell types significantly induced tubule formation (*) as determined by Student's t-test (p < 0.05). Drug treatments that significantly reversed the effects of conditioned medium are indicated (**) as determined by ANOVA and Student's t-test (p < 0.05). b) Monolayers of HUVEC cells were allowed to reach a minimal TEER plateau and subsequently incubated with conditioned medium from UMCRC2 (left panel) or 786-O cells (right panel) in the presence or absence of 17-AAG, EC154, or LBH589. Conditioned medium from HUVEC cells was used as a negative control. Impedance was measured at 5 min intervals, normalized to levels just prior to the addition of effectors, and presented relative to untreated control. The traces shown represent an average of two replicates per condition.