Figure 6. VEGFA₁₆₅a has a stronger effect on the expression of tight-junction-protein genes in primary HRMECs than VEGFA₁₆₅b. Relative gene expression measured with quantitative PCR (qPCR). A: Expression of the CLDN5 gene at 1, 3, 6, and 24 h after treatment with VEGFA₁₆₅. Confluent human retinal microvascular endothelial cells (HRMECs) were treated with a saturating high dose of VEGFA₁₆₅a or VEGFA₁₆₅b (100 ng/ml, 5,300 pM). B: Expression of the CLDN5 gene comparing VEGFA₁₆₅a and VEGFA₁₆₅b at low, intermediate, and high doses measured after 6 h. C: Expression of the OCLN gene at 1, 3, 6, and 24 h after treatment with VEGFA₁₆₅. D: Expression of the OCLN gene comparing VEGFA₁₆₅a and VEGFA₁₆₅b at low, intermediate, and high doses measured after 6 h. (Triplicate assays, error bars show standard deviation. ANOVA (ANOVA): compared to media control *p<0.05, **p<0.01; compared to VEGFA₁₆₅a ^†† p<0.01).