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. 2015 Dec 2;17(1):53–65. doi: 10.1111/tra.12341

Figure 2.

TRA-12341-FIG-0002-b

USP8 depletion promotes generation of a novel 120 kDa VEGFR2 proteolytic cleavage fragment. Endothelial cells transfected with non‐targeting or USP8 siRNA were treated with 25 ng/mL VEGF‐A (A) or 20 µg/mL CHX and 25 ng/mL VEGF‐A (B), lysed and immunoblotted with antibodies against VEGFR2. Quantification of 120 kDa (C) or 160 kDa (D) VEGFR2 fragment levels in endothelial cells transfected with non‐targeting or USP8 siRNA and treated with 20 µg/mL CHX and 25 ng/mL VEGF‐A. E) VEGFR1 or VEGFR2 were immunoprecipitated from endothelial cells transfected with non‐targeting or USP8 siRNA and immunoblotted with antibodies to the extracellular and cytoplasmic domains of VEGFR2 (F) or run alongside marker ladders to confirm band size. Numbered arrowheads denote the 160 kDa VEGFR2 fragment 1 and the novel 120 kDa VEGFR2 fragment 2. Error bars denote ±SEM (n ≥ 3); p < 0.05 (*), p < 0.01 (**).