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Figure 4


Fig. 4. Tbdn/Ard1 knockdown in endothelial cells is associated with increased cellular permeability. (A) RF/6A parental endothelial cells (Par), Tbdn-knockdown clones AS-TBDN (ASTB#1 and #2) and control clones (Ctr#1 and #2) analyzed by western blot for Tbdn and Ard1 expression (top and middle panels, respectively). Blots were re-probed and analyzed for tubulin as loading control (bottom panel). (A) Representative experiment; (B) the average of Tbdn and Ard1 levels ± s.e.m. are shown. (C) FITC-albumin transit across monolayers of RF/6A knocked-down for Tbdn expression (ASTB#1 and #2) or controls (Par, CTR#1 and #2) expressed as a percentage of control parental cells at 30 minutes. Significantly higher percentages of FITC-albumin transit are observed in the two AS-TBDN clones compared with parental cells and the two negative-control clones. (D) Time-course of FITC-albumin transit across cellular monolayers of Tbdn-knockdown RF/6A cells (ASTB#1; black square) as compared with parental cells (black diamond) and control cells (CTR#1; white triangle) expressed as percentage of arbitrary units. Data shown in C and D are expressed as mean ± s.e.m. of at least four duplicate experiments in each group.





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