SVEC4-10 cells (American Type Culture Collection, USA) were used for LEC in vitro experiments (7). Cell Counting Kit-8 (CCK-8) assay: VEGF-C or VEGF-D (0, 1, 10, and 100 ng/μl) and FGF-2 (0, 1, 10, and 20 ng/μl) were added to promote cell proliferation. sVEGFR3-FC (100 ng/μl) or sLYVE-1-FC (250 ng/μl) was added to inhibit proliferation. sFC at the same concentration was used as control. Forty-eight hours after stimulation, SVEC4-10 cells in 96-well plates were treated with 10 μl of CCK8 and 90 μl of Iscove’s modified Dulbecco’s medium (IMDM) (Gibco, USA) and then incubated at 37°C with 5% CO2. Optical density (OD) values were measured using a microplate reader (BioTek, USA). 5-ethynyl-2′-deoxyuridine (EdU) assay: 48 hours after stimulation, SVEC4-10 cells in 96-well plates were incubated with 100 μl of 50 μmol EdU for 2 hours and then immobilized with 4% polyoxymethylene. EdU staining was imaged with Cell-Light EdU Apollo 567 In Vitro Imaging Kit (C10310-1, RiboBio Co., Guangzhou, China) according to the manufacturer’s instructions. Nuclei were stained with Hoechst 33342. Apoptosis assay: 24 hours following stimulation with either sVEGFR3-FC (100 ng/μl) or sLYVE-1-FC (250 ng/μl), an Annexin V–FITC (AV)/propidium iodide (PI) assay was performed to analyze apoptosis (BD Biosciences, USA). Flow cytometry was then used to detect AV- and PI-positive cells.

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