Fig. 5

The effect of miR-548k on VEGFR3 phosphorylation. a, Immunoprecipitation and Western blot assays were used to examine the interaction of ADAMTS1 and VEGFC in miR-548k overexpression cells (KYSE30-Lenti-miR-548k) and control cells. Total cell lysates (TCL) of miR-548k overexpression cells and control cells were immunoprecipitated with the antibody against the indicated proteins. Immunocomplexes were then immunoblotted using antibodies against the indicated proteins. TCL were also immunoblotted using antibodies against the indicated proteins. b, The secretory VEGFC was examined in supernatants of miR-548k overexpression cells and control cells. c, The secretory VEGFC was examined in supernatants of ADAMTS1 and or miR-548k overexpression cells and control cells. d, The phosphorylation level of VEGFR3 in HDLEC cells. HDLEC cells were cultured in conditioned media of miR-548k overexpression cells and control cells. Cells were immunoprecipitated with the antibody against VEGFR3. Immunocomplexes were then immunoblotted using antibodies against the indicated proteins. e, The phosphorylation level of VEGFR3 in HDLEC cells. HDLEC cells were cultured in conditioned media of ADAMTS1 and/or miR-548k overexpression cells and control cells. Cells were immunoprecipitated with the antibody against VEGFR3. Immunocomplexes were then immunoblotted using antibodies against the indicated proteins. f, Illustrative model showing the proposed mechanism by which miR-548k promotes cell lymphangiogenesis and lymphatic metastasis in ESCC via suppressing ADAMTS1 expression