Fig 1: The silence of EGFL6 reduced the growth of NPC in nude mice. A, Representative examples of nude mice subcutaneous injection of CNE2 cells. B, Tumor volumes were evaluated every two days. C, The final tumor volume of sacrificed mice at day 12 was obtained. Mean ± SEM, *P < 0.05
Fig 2: The expression of EGFL6 in nasopharyngeal carcinoma (NPC). A, High expression of EGFL6 was observed in NPC tissues compared with non-cancerous nasopharyngeal (N) tissues by immunohistochemistry analysis. B, High level of EGFL6 in NPC serum compared with N serum. **P < 0.01. C, The standard curve analysis of EGFL6 in serum
Fig 3: Deletion of EGFL6 in osteoblasts leads to impaired bone repair characterized by reduced angiogenesis. (A) Schematic illustration of mono-cortical bone defect model. (B) Representative three-dimensional reconstructed micro-CT images of bone repair in EGFL6OCN and EGFL6fl/Y mice 1 week after surgical procedure, and bone histomorphometric analysis including hematoxylin-eosin (HE), picrosirius red (PSR), Masson trichrome staining of bone defect region. CB, cortical bone. Scale bar = 200 μm (C) Quantification of newly formed bone in defect region by micro-CT scanning and Masson trichrome staining (n = 5 per group). BV/TV, bone volume per tissue volume; BS/TS, bone surface and tissue surface. (D) Representative micro-CT and bone histomorphometric images of bone defect at week 2. Scale bar = 200 μm.(E) Quantification of the newly formed bone in defect region at week 2 by micro-CT scaning (WT, n = 11; cKO, n = 10) and Masson trichrome staining (n = 5 per group). (F) Representative confocal images of type H vessels stained for CD31 (red) and EMCN (green) in bone defect region of EGFL6OCN and EGFL6fl/Y mice at week 2. (G) Quantification of EMCN+ and CD31+ area in (F) (n = 5 per group). White dashed lines indicate the edge of bone tissue. All bar graphs are presented as mean ± SD. *P < 0.05, **P < 0.01 relative to the WT group. Differences are analyzed using Student's t-test.
Fig 4: Effect of EGFL6 knockout on tumor growth and angiogenesis in the ovarian cancer nude mouse model. Ovarian cancer cells with EGFL6 knockout (E10 and G11, 6 × 106 cells in 100 µl PBS) were injected subcutaneously in the right shoulder of BALB/c nude mice (n = 8). The SKOV3 cells were the control. (A) Tumors stripped from the nude mice in different groups. (B) The curve of tumor weight changes in the nude mice in different groups. (C) Growth curve of tumors in different groups. (D) Immunohistochemical assay of tumor blood and lymphatic vasculature and angiogenic factors. The blood vascular network was stained with anti-CD31 antibody. The lymphatic vasculature was stained with anti-LYVE1 antibody. The angiogenic factors were assayed with the antibodies against EGFL6, FGF2, PDGFB, and VEGFA. *p < 0.05 and **p < 0.01 were considered statistically significant.
Fig 5: Kaplan-Meier actuarial analysis of EGFL6 expression with respect to overall survival of (A) all patients and (B) patients younger than 69 years of age. EGFL6, epidermal growth factor-like domain 6.
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