American Society of Hematology

Figure 3.

$Figure 3. VEGF stimulation redirects KDR to the late endosomal compartment, but only a fraction of receptor undergoes subsequent lysosomal degradation. HUVECs were stimulated with 100 ng/mL VEGF for 0 hours (A,D), 1 hour (B,E), or 24 hours (C-D) and then fixed and stained (green) for endogenous EEA1 (A-C) or CD63 (D-F). All cells were costained for endogenous KDR (red). Nuclei are stained with DAPI (blue). Inset panels show magnified portions of each image, as indicated (dashed squares). Bar represents 10 μm. (G) The percentage of KDR+ vesicles that colocalized with EEA1+ vesicles or CD63+ vesicles at the 3 time points was quantified (mean ± SD; n ≥ 16). Degradation of KDR over the same time course of VEGF stimulation was determined by Western blotting of HUVEC lysates (H). The lower portion of the Western blot was probed for tubulin to confirm equal loading between samples. (I) Densitometric quantification of the degradation time course experiments (mean ± SD; n = 4).$

VEGF stimulation redirects KDR to the late endosomal compartment, but only a fraction of receptor undergoes subsequent lysosomal degradation. HUVECs were stimulated with 100 ng/mL VEGF for 0 hours (A,D), 1 hour (B,E), or 24 hours (C-D) and then fixed and stained (green) for endogenous EEA1 (A-C) or CD63 (D-F). All cells were costained for endogenous KDR (red). Nuclei are stained with DAPI (blue). Inset panels show magnified portions of each image, as indicated (dashed squares). Bar represents 10 μm. (G) The percentage of KDR⁺ vesicles that colocalized with EEA1⁺ vesicles or CD63⁺ vesicles at the 3 time points was quantified (mean ± SD; n ≥ 16). Degradation of KDR over the same time course of VEGF stimulation was determined by Western blotting of HUVEC lysates (H). The lower portion of the Western blot was probed for tubulin to confirm equal loading between samples. (I) Densitometric quantification of the degradation time course experiments (mean ± SD; n = 4).

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