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. 2018 May 18;32(11):5793–5812. doi: 10.1096/fj.201700818RRR

Figure 6.

Figure 6

CREB3 knockdown restores endothelial barrier function. A) Permeability of control (siCtrl) or CREB3-silenced (siCREB3) HUVEC monolayer was determined using Transwell chamber system after 48 h of cell incubation with DMSO or forskolin. FITC-dextran (70 kDa, 1 mg/ml) was added to upper chamber, and fluorescence signal in lower compartment was measured by fluorophotometer at different time points. Empty chambers without cells were used as reference (100% dextran leakage) for percentage calculation. Ns, not significant.**P < 0.01, ****P < 0.0001 (3-way ANOVA). B) Confluent monolayer of control (siCtrl) or CREB3-silenced (siCREB3) HUVEC was treated with DMSO or forskolin for 48 h and subsequently stained for VE-cadherin. Quantification of immunostaining was performed by measuring integrated fluorescence intensity of 3 different areas from at least 4 micrographs for each condition. Scale bar, 100 µm. Ns, not significant. *P < 0.05 (2-way ANOVA). C) VE-cadherin expression in CREB3-silenced ECs in DMSO control or forskolin-treated culture was analyzed by Western blot.