FIGURE 5.

PDCL3 protects VEGFR-2 from proteolytic degradation. PAE cells coexpressing VEGFR-2 with an empty vector or with PDCL3 were lysed, and whole cell lysates were divided into two groups. One of the lysate groups was denatured by boiling at 95 °C, and both groups were subjected to immunoprecipitation (IP) using anti-VEGFR-2 antibody. Immunoprecipitated proteins were analyzed by Western blot analysis using anti-c-myc antibody (A), anti-HSP70 antibody (B), and anti-PLCγ1 antibody (C). The whole cell lysates from the same groups were also blotted with anti-c-myc antibody, anti-Hsp70 antibody, and anti-PLCγ1 antibody. Cell lysates from PAE cells coexpressing VEGFR-2 with an empty vector or PDCL3 were treated with trypsin (25 ng/50 μl of cell lysate) or vehicle, followed by Western blot analysis using anti-VEGFR-2 antibody (E), anti-IQGAP1 antibody as a loading control (F), and anti-c-myc antibody for PDCL3 (G).