Fig 1: Comparison of biological functions of LECs. a Determination of in vitro LECs tube formation ability by Calcein AM (green) in vitro (scale bar, 50 µm). b Endothelial cell phagocytosis assay (DiI-labeled, red; DAPI, blue) of endothelial cell function using isolated LECs by LYVE-1 Microbeads and Fe3O4@KCTS-LECs-Double antibody (scale bar, 100 µm)
Fig 2: Analysis of the purity of lymphatic endothelial cells. a Fluorescence micrographs double-color imaging of LYVE-1 (red) and podoplanin (green)/nuclei [4,6-diamidino-2-phenylindole (DAPI, blue) in the isolated human colorectal cancer LECs sorted using LYVE-1 Microbeads and Fe3O4@KCTS-LECs-double antibody (scale bar, 100 µm). b The co-expression of LYVE-1 (APC) and podoplanin (FITC) as revealed by flow cytometric detection. c Results of flow cytometry, *P < 0.05
Fig 3: CCBE1 and LYVE1 expression in patients with lung cancer and healthy controls. (A) Reverse transcription-polymerase chain reaction analysis of CCBE1 mRNA expression levels in lung tissue samples. Western blot analysis of (B) CCBE1 and (C) LYVE-1 protein expression levels in lung tissue samples. Lanes 1–3 represent samples from normal tissue; lanes 4 and 5 represent samples from patients with cancer and LNM; lanes 6–8 represent samples from patients with cancer without LNM. Data are expressed as the mean ± standard deviation (n=5 in each group). *P<0.05 vs. normal group; #P<0.05 vs. LNM group. CCBE1, collagen and calcium-binding epidermal growth factor domain-containing protein 1; LYVE1, lymphatic vessel endothelial hyaluronan receptor 1; LNM, lymph node metastasis.
Fig 4: Expression of lymphatic vessels in colorectal cancer tissues. a Dilated vessels in the tissue sections were positive for LYVE-1 on endothelial cell membrane by immunohistochemistry (scale bar, 100 µm). On the right is a magnified picture. b Dilated vessels in the tissue sections were positive for LYVE-1 and podoplanin on endothelial cell membrane as revealed by immunofluorescence (scale bar, 50 µm)
Fig 5: LYVE1 expression in patients with lung cancer and healthy controls. Representative cytoplasmic LYVE1 staining from a total of 8 patients is demonstrated. (A) Tissues isolated from control patients who underwent pulmonary bullae resection. (B) Tissues isolated from patients with lung cancer without LNM. (C) Tissues isolated from patients with lung cancer accompanied by LNM. Black arrows indicate LYVE1 staining patterns. Photomicrographs were captured under ×200 magnification. LYVE, lymphatic vessel endothelial hyaluronan receptor; LNM, lymph node metastasis.
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