CD36 and SHP-1 are required for TSR inhibition of MVEC tubelike structure formation. (A) MVEC were transfected with CD36 or control siRNA as in Figure 3 and then treated with 10 nmol/L TSR in low-serum medium for 4 hours before being transferred onto Matrigel-coated tissue culture wells. Cells were then exposed to 50 ng/mL VEGF for 6 hours, fixed with 4% paraformaldehyde, and then stained with 30 nmol/L fluorescence-labeled Phalloidin for 1 hour. Images from 4 randomly chosen areas were obtained with a fluorescence microscope, and the number of branches and average branch length of tubelike structures were quantified using NIH ImageJ software. The scale bar represents 200 μm. (B) MVEC were transfected with control siRNA and treated with TSR as in panel A. Cells were then exposed to 50 ng/mL VEGF for 6 hours in the presence of 100 μmol/L SHP inhibitor NSC-87877 or vehicle control. The number of branches and the average branch length of tubelike structures were analyzed as in panel A.