Fig. 3.

Agrin 000 and agrin 748 do not increase expression levels of VE-cadherin, β-catenin and ZO-1 in bEnd5 cells. a, c, e Quantitative reverse transcription plus the polymerase chain reaction was performed to analyze VE-cadherin, β-catenin, and ZO-1 mRNA levels in bEnd5 cells cultured for 24 h on agrin 000, agrin 748, or under control conditions. Relative expression (RQ) values were normalized to the control condition for every independent experiment. Bars represent the mean of three independent experiments (± SEM). s16 ribosomal protein mRNA served as endogenous control. b, d, f bEnd5 total VE-cadherin, β-catenin, and ZO-1 protein amounts were determined by Western blot analysis after 48 h in culture. Protein levels were normalized to the endogenous control protein β-actin. Values were normalized to the control condition for every independent experiment. Bars represent the mean of three (VE-cadherin, β-catenin) and six (ZO-1) independent experiments (± SEM). *P < 0.05 (n.s. not significant)