Figure 3 Association of CD36 with... | American Society of Hematology

Figure 3

$Figure 3. Association of CD36 with VEGFR-2, Syk, and integrin β1 subunit in the lungs of wild-type and TSP-1--null mice. (A) Brij 99 lysates from TSP-1–null and wild-type lung tissues were immunoprecipitated with nonimmune IgG (lane 1) or anti-CD36 Ab (lane 2) and Western blotted with anti-β1 integrin Ab. (B) Top: Triton X-100 lysates from TSP-1–null and wild-type lung tissues were immunoprecipitated with nonimmune IgG (lane 1) or an anti-Syk Ab (lane 2) and Western blotted with an anti-CD36 Ab. Middle: In parallel experiments, tissue extracts were immunoprecipitated with anti-CD36 Ab (lane 2) and nonimmune IgG (lane 1) and Western blotted with anti-Syk and anti-CD36 Abs, as indicated. Bottom: The high-density fractions of sucrose gradients prepared with Triton X-100 extracts of TSP-1-null and wild-type lung tissues (supplemental Figure 6B) were immunoprecipitated with nonimmune IgG (lane 1) or anti-CD36 Ab (lane 2) and Western blotted with an anti-Syk Ab. (C) Top: Brij 99 lysates from TSP-1–null and wild-type lung tissues were immunoprecipitated with nonimmune IgG (lane 1) or anti-CD36 Ab (lane 2) and Western blotted with anti-VEGFR2 Ab. In this experiment, the total lysates were used as loading control (middle panel). Bottom: Brij 99 lysates from TSP-1–null and wild-type lung tissues were immunoprecipitated with nonimmune IgG (lane 1) or anti-VEGFR-2 Ab (lane 2) and Western blotted with anti-CD36 Ab. The immunoprecipitates were also Western blotted with an anti-VEGFR-2 Ab to establish that comparable amounts of VEGFR-2 were immunoprecipitated from the TSP-1–null and wild-type tissue. (D) Top: Brij 99 lysates from TSP-1–null and wild-type lung tissue were immunoprecipitated with nonimmune IgG (lane 1) or anti-Syk Ab (lane 2) and Western blotted with anti-VEGFR-2 or anti-Syk Abs, as indicated. Bottom: Brij 99 lysates from TSP-1–null and wild-type lung tissue were immunoprecipitated with nonimmune IgG (lane 1) or anti-VEGFR-2 Ab (lane 2) and Western blotted with anti-Syk and anti-VEGFR-2 Abs. In these experiments, either total lysates or immunoprecipitants were also Western blotted to establish that comparable amount of proteins were detected or immunoprecipitated from TSP-1–null and wild-type tissues.$

Association of CD36 with VEGFR-2, Syk, and integrin β1 subunit in the lungs of wild-type and TSP-1--null mice. (A) Brij 99 lysates from TSP-1–null and wild-type lung tissues were immunoprecipitated with nonimmune IgG (lane 1) or anti-CD36 Ab (lane 2) and Western blotted with anti-β1 integrin Ab. (B) Top: Triton X-100 lysates from TSP-1–null and wild-type lung tissues were immunoprecipitated with nonimmune IgG (lane 1) or an anti-Syk Ab (lane 2) and Western blotted with an anti-CD36 Ab. Middle: In parallel experiments, tissue extracts were immunoprecipitated with anti-CD36 Ab (lane 2) and nonimmune IgG (lane 1) and Western blotted with anti-Syk and anti-CD36 Abs, as indicated. Bottom: The high-density fractions of sucrose gradients prepared with Triton X-100 extracts of TSP-1-null and wild-type lung tissues (supplemental Figure 6B) were immunoprecipitated with nonimmune IgG (lane 1) or anti-CD36 Ab (lane 2) and Western blotted with an anti-Syk Ab. (C) Top: Brij 99 lysates from TSP-1–null and wild-type lung tissues were immunoprecipitated with nonimmune IgG (lane 1) or anti-CD36 Ab (lane 2) and Western blotted with anti-VEGFR2 Ab. In this experiment, the total lysates were used as loading control (middle panel). Bottom: Brij 99 lysates from TSP-1–null and wild-type lung tissues were immunoprecipitated with nonimmune IgG (lane 1) or anti-VEGFR-2 Ab (lane 2) and Western blotted with anti-CD36 Ab. The immunoprecipitates were also Western blotted with an anti-VEGFR-2 Ab to establish that comparable amounts of VEGFR-2 were immunoprecipitated from the TSP-1–null and wild-type tissue. (D) Top: Brij 99 lysates from TSP-1–null and wild-type lung tissue were immunoprecipitated with nonimmune IgG (lane 1) or anti-Syk Ab (lane 2) and Western blotted with anti-VEGFR-2 or anti-Syk Abs, as indicated. Bottom: Brij 99 lysates from TSP-1–null and wild-type lung tissue were immunoprecipitated with nonimmune IgG (lane 1) or anti-VEGFR-2 Ab (lane 2) and Western blotted with anti-Syk and anti-VEGFR-2 Abs. In these experiments, either total lysates or immunoprecipitants were also Western blotted to establish that comparable amount of proteins were detected or immunoprecipitated from TSP-1–null and wild-type tissues.

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