Skip to main content
. 2017 Nov 20;293(2):717–730. doi: 10.1074/jbc.M117.815381

Figure 3.

Figure 3.

8-CPT-AM blocks and reverses permeability induced by VEGF and TNF-α. A, BREC were stimulated with VEGF (50 ng/ml) or/and 8-CPT-AM (1 μm). Pre−, BREC were stimulated with 8-CPT-AM 30 min before VEGF treatment. Post−, BREC were treated with 8-CPT-AM 30 min after VEGF. 70 kDa RITC-dextran was added 30 min after the last treatment. Average Po values for control and VEGF were 9.7 × 10−7 and 1.7 × 10−6 (cm/s), respectively, n ≥ 6. B, BREC were pretreated for 30 min before stimulated with TNF-α (10 ng/ml) and 70 kDa RITC-dextran was added 1 h later. Average Po values for control and TNF-α were 1.1 × 10−6 and 1.7 × 10−6 (cm/s), n ≥ 12. C, BREC were stimulated with TNF-α (10 ng/ml) for 1 h followed by VEGF for 30 min. Pre−, 8-CPT-AM (1 μm) was added 30 min before TNF-α + VEGF or Post−, 8-CPT-AM (1 μm) was added 1.5 h after TNF-α + VEGF. 70-kDa RITC-dextran was added 30 min after the last treatment. Average Po values for control were 7.5 × 10−7 and for TNF-α + VEGF were 1.9 × 10−6 (cm/s), n ≥ 15. Results are expressed as the mean relative to the control ± S.D. One-way analysis of variance: ##, p < 0.005; ###, p < 0.001; ####, p < 0.0001 compared with control. ****, p < 0.0001 compared with VEGF, TNF-α, or VEGF + TNF-α in A, B, and C, respectively. D, BREC were seeded on 8W10E+ arrays and TEER was measured every hour on the ECIS Zϴ instrument. 8-CPT-AM (2 μm) added 30 min prior to VEGF increases resistance and prevents VEGF-induced TEER loss for up to 6 h. 8-CPT-AM post-VEGF restores endothelial barrier after VEGF-induced ion permeability. Data represents the mean ± S.E. with analysis by two-way analysis of variance Bonferroni post hoc test: ****, p < 0.0001 compared with VEGF, n = 3/group.