Purchase this article with an account.
Jin-Hong Chang, Kyu-Yeon Han, Dimitri T. Azar; Involvement of Lysosomal Proteases in VEGF-C Down-Regulation of VEGFR-3. Invest. Ophthalmol. Vis. Sci. 2012;53(14):2394.
Download citation file:
© ARVO (1962-2015); The Authors (2016-present)
To characterize the involvement of Lysosomal Proteases in VEGF-C Down-Regulation of VEGFR-3
Lymphatic endothelial cells were stimulated with vascular endothelial growth factor (VEGF-C, -D, C156S, and VEGF-A) at various (vague) time-points. Unstimulated and stimulated cells were lysed with RIPA buffer and cellular lysates were immunoblotted with anti-VEGFR-3, phosphor-tyrosine, Erk, jnk, and p38 antibodies. Lymphatic cells were also stimulated with VEGF-C in the presence of a VEGF-C trap, Notch activator, inhibitor, VEGF Receptor-3 (VEGFR-3) kinase inhibitor, LPE (leupeptin, pepstatin, and E64 inhibitor), gamma-secretase inhibitor, or autophagy inhibitor Cellular lysates were immunobloted with anti-VEGFR-3 antibodies.
VEGF-C induces down-regulation of VEGFR-3, which is important for lymphangiogenesis, but incompletely understood. Here, we demonstrate that VEGF-C, -D, and C156S, but not VEGF-A, down-regulate VEGFR-3. VEGF-C stimulated VEGFR-3 tyrosyl phosphorylation and transient phosphorylation of ERK, p38, and c-Jun N-terminal kinases in lymphatic endothelial cells. VEGF-C-induced VEGFR-3 down-regulation was blocked by a VEGF-C trap, VEGFR-3 kinase inhibitor, and LPE inhibitor, but was unaffected by Notch 1 activator and gamma-secretase inhibitors.
Our findings indicate that VEGF-C down-regulates VEGFR-3 in lymphatic endothelial cells through VEGFR-3 kinase activation and, in part, via the lysosomal degradation pathway.
This PDF is available to Subscribers Only