FIGURE 7.

KGF and HGF protect the endothelium from H₂O₂-induced barrier dysfunction. (A,B) HPAEC grown on gold electrodes of ECIS arrays were exposed to the original growth media (B-red), or growth media supplemented with KGF (B-orange), HGF (B-blue), KGF + HGF (B-green), or VEGF concentrations detected in ASC-CM conditioned for 48 h (see Figure 6). Media supplemented with factors were removed after 48 h of pretreatment, and HPAEC were challenged with 250 μM H₂O₂. For (A), TER values were pooled from three independent experiments, with at least two parallel recordings each, and calculated as fold decrease in TER 30 min after H₂O₂ addition. (C) HPAEC grown on gold electrodes of ECIS arrays were exposed to the original growth media, or ASC-CM generated by ASC treated with scrambled siRNA, or HGF siRNA. Media were removed after 48 h of pretreatment, and HPAEC were challenged with 250 μM H₂O₂. Inset shows the concentration of HGF in ASC-CM collected from scrambled RNA-treated and HGF siRNA-treated ASC. (D) HPAEC grown on gold electrodes of ECIS arrays were exposed to the original growth media, or ASC-CM pre-incubated with control IgG or KGF neutralizing antibody. Media were removed after 48 h of pretreatment, and HPAEC were challenged with 250 μM H₂O₂. (A) ^∗Differences detected with one-way ANOVA with Tukey post hoc (p < 0.05). (B) Repeated measurements ANOVA detected differences between responses of HPAEC exposed to growth media supplemented with either KGF or HGF (^∗), and the original growth media. Differences were also detected between responses of HPAEC exposed to media supplemented with KGF and KGF + HGF (#). (C,D) ^∗ Repeated measurements ANOVA detected differences between responses of HPAEC exposed to unmanipulated ASC-CM or ablated ASC-CM. (C, inset) ^∗ Differences were detected by t-test (p < 0.05).