American Society of Hematology

Figure 3

$Figure 3. Ad-siCav-1 causes loss and redistribution of AJ-associated proteins. BMECs were exposed to Ad-siCav-1 or Ad-siLuc (control) at 500 ifu per cell for 96 hours and then subjected to analysis. (A) Immunofluorescent staining for occludin and ZO-1 showing reduced and fragmented staining patterns following Ad-siCav-1 exposure. Scale bar represents 50 μm. Western blotting (B) and corresponding quantification (C) showing loss of VE-cadherin and β-catenin in response to Ad-siCav-1. Western blotting (D) and corresponding quantification (E) revealing a significant shift in the association of both VE-cadherin and β-catenin from the cytoskeletal framework (CSK) to the soluble (SOL) fraction of BMECs (increased SOL/CSK ratio) as a result of Ad-siCav-1 treatment. Because equal amounts of total protein were analyzed in panel B, while equal aliquots (cellular equivalents) were analyzed in panel D, protein intensities between these 2 figures are not comparable. Blots are representative of 3 experiments. Data are presented as mean ± SE of all 3 experiments. *P < .01.$

Ad-siCav-1 causes loss and redistribution of AJ-associated proteins. BMECs were exposed to Ad-siCav-1 or Ad-siLuc (control) at 500 ifu per cell for 96 hours and then subjected to analysis. (A) Immunofluorescent staining for occludin and ZO-1 showing reduced and fragmented staining patterns following Ad-siCav-1 exposure. Scale bar represents 50 μm. Western blotting (B) and corresponding quantification (C) showing loss of VE-cadherin and β-catenin in response to Ad-siCav-1. Western blotting (D) and corresponding quantification (E) revealing a significant shift in the association of both VE-cadherin and β-catenin from the cytoskeletal framework (CSK) to the soluble (SOL) fraction of BMECs (increased SOL/CSK ratio) as a result of Ad-siCav-1 treatment. Because equal amounts of total protein were analyzed in panel B, while equal aliquots (cellular equivalents) were analyzed in panel D, protein intensities between these 2 figures are not comparable. Blots are representative of 3 experiments. Data are presented as mean ± SE of all 3 experiments. *P < .01.

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