Figure 7

The effects of DIF-1 on the proteolysis and the proteosynthesis of VEGFR-2. (A) DIF-1 did not affect the degradation speed of VEGFR-2. HUVECs were incubated with or without DIF-1 (30 μM) for the given periods in the presence of cycloheximide (10 μM). Protein samples were subjected to Western blot analysis for VEGFR-2. The membrane was re-probed with anti-GAPDH antibody. The levels of protein bands were quantified and are shown as percentage of the control level at time 0. Values are mean ± SE of three independent experiments. (B) DIF-1 attenuated the protein synthesis of VEGFR-2. After HUVECs were pretreated with cycloheximide (10 μM) for 3 h, the medium was changed to fresh medium and cells were incubated with or without DIF-1 (30 μM) for the given periods. Protein samples were subjected to Western blot analysis for VEGFR-2. The membrane was re-probed with anti-GAPDH antibody. The levels of protein bands were quantified and are shown as percentage of the control cells which were not treated with cycloheximide (lane 1). Values are mean ± SE of three independent experiments. The asterisk indicates *P < 0.001 versus control.