Fig. 4

Tight junction protein expression is not altered by pUR4. After being pretreated with 1000 nM scrambled peptide or 1000 nM pUR4 for 16 h, bEND.3 cells were stimulated with 20 ng/mL TNF-α for the indicated durations. The protein expression of ZO-1 (a), claudin-5 (c), and occludin (e) in bEND.3 cells 24 h after TNF-α treatment was evaluated through immunoblotting; the quantitative analysis results for of ZO-1, claudin-5, and occludin are normalized to GAPDH or α-tubulin (n = 3). ZO-1 (b), claudin-5 (d), and occludin (f) transcript expression in bEND.3 cells treated with TNF-α for 16 and 36 h was evaluated using quantitative real-time PCR (n = 3). Data are presented as means ± standard deviations. **P < 0.01 and ****P < 0.0001, one-way ANOVA followed by Tukey’s multiple comparison test