Figure 8

Endocytic recycling of vascular endothelial growth factor receptor 2 (VEGFR2) post-shock wave (SW) treatment. (A) VEGFR2, Na/K ATPase (marker of membrane protein), Lamin B (marker of nucleus protein) and actin in protein fraction of HUVECs (including membrane, cytosol and nucleus parts) at the end of the first 4 and 28 h post-SW treatment compared with control (CON) assessed by Western blot. (B) The workflow noted three time points for SW, drug treatment and tube formation. Protein expressions of VEGFR2 (with short and long exposure time), Na/K ATPase and actin in HUVECs (fractionized into membrane and cytosol parts) assessed by Western blot 26 h following DMSO, cycloheximide (CHX) and chloroquine (CHQ) treatment with and without SW administration. (C) Tube formation assay for HUVECs 26 h post being treated with cycloheximide (CHX) and chloroquine (CHQ, 5 & 20 µmol/L) in basal medium (BM, without FBS and growth factors) with and without SW treatment. (D) Parameters of tube formation assay, including tube length, branch point and the number of loops (n = 6 in each group). (E) Tube formation assay for HUVECs 26 h after being treated with DMSO, cycloheximide (CHX) and high concentration of chloroquine (CHQ, 20 µmol/L) in complete medium (CM, that is, contained FBS and growth factors). (F) Parameters of tube formation assay, including tube length, branch point and the number of loops (n = 5 in each group). Data shown as mean ± S.D. ***P < 0.0001 determined by Student t test.