Fig. 6
ANGPTL4 knockdown loses its tumor-suppressive effect in VEGFR2 Y949F/Y949F mice. a Experimental strategy. MISIIR-TAg mice (TAg mice) were generated, the sex of the pups was determined, and tail tips were acquired from female mice for genotyping. On postnatal day (pnd) 28, with control lentiviral vectors (LVs) or LVs encoding siRNA targeting ANGPTL4 were injected into the ovaries of mice. On pnd 56, mice were sacrificed for further analysis. b Ovarian tumor volumes in control (Ctl)-injected and siANGPTL4 (siANG)-injected mice on pnd 56; n = 10 mice per group. c Distribution of tumor diameters (1–5 mm and > 5 mm) on pnd 56; n = 10 mice per group. d Experimental strategy. VEGFR2Y949F/Y949F mice were crossed with MISIIR-TAg mice on a C57BL/6 background to obtain TAg-WT (WT) and TAg-VEGFR2Y949F/Y949F (VEGFR2Y949F/Y949F or Y949F) homozygous mice with induced spontaneous epithelial ovarian carcinoma and/or continuous inactivation of VEGFR2 at tyrosine 949. On pnd 28, LVs encoding siRNA targeting ANGPTL4 were injected into the ovaries of mice. On pnd 56, mice were sacrificed for further analysis. e WT and VEGFR2Y949F/Y949F ovarian tumor volumes on pnd 56; n = 20–23 mice per genotype. f Distribution of tumor diameters (1–5 mm and > 5 mm) on pnd 56; n = 20–23 mice per genotype. g EdU-positive (green) cell/simian virus 40 large T antigen (SV40, red)-positive tumor cell ratio in ovarian tissue; n = 7 tumors per genotype. Scale bar: 30 μm. h Immunofluorescence staining and statistical graph of CD31 expression in WT and VEGFR2Y949F/Y949F ovarian tumors; n = 6 tumors per genotype. Scale bar: 50 μm. i Immunofluorescence staining and statistical graph of VE-cadherin expression in Ctl and siANG tumors or WT and VEGFR2Y949F/Y949F tumors; n = 4–6 tumors per group. Scale bar: 50 μm. j Western blot of VE-cadherin and GAPDH in Ctl and siANG tumors or WT and VEGFR2Y949F/Y949F tumors; n = 4 tumors per group. k Western blot of slug, snail, and GAPDH in Ctl and siANG tumors or WT and VEGFR2Y949F/Y949F tumors; n = 4 tumors per group. *p < 0.05, **p < 0.01, ***p < 0.001 for the indicated comparisons. The data are presented as the mean ± SEM values