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. Author manuscript; available in PMC: 2020 Jul 15.
Published in final edited form as: Nature. 2020 Jan 15;577(7792):689–694. doi: 10.1038/s41586-019-1912-x

Extended Data Fig 4. VEGF-C signals specifically in lymphatic endothelial cells in the meninges and dCLNs and provides survival benefits in an administration timepoint dependent manner.

Extended Data Fig 4.

(a) Gating strategy for lymphatic endothelial cells (LECs) and blood endothelial cells (BECs). (b) Concatenated images of LECs and BECs from meninges and lymph node depicting AKT-phosphorylation intensity. Experiment was repeated independently with similar results. (c) Quantification of AKT(pS473) positive population and MFI within LECs and BECs in the meninges and dCLNs (meninges; WT, n = 5; AAV-VEGF-C, tumor + Luc-mRNA, tumor + VEGF-C-mRNA, n = 8) (lymph nodes; WT, n = 5; AAV-VEGF-C, n = 8; tumor + Luc-mRNA, n = 7; tumor + VEGF-C-mRNA, n = 8). (d) Fluorescent microscope images of dCLN after VEGF-C-mRNA treatment in tumor bearing mice (CD31, red; LYVE1, green; DAPI, blue). (e) Fluorescent microscope images of meninges after VEGF-C-mRNA treatment in tumor bearing mice (CD31, red; LYVE1, green; DAPI, blue). Experiment was repeated independently with similar results. f-h Mice were treated with AAV-VEGF-C or VEGF-C-mRNA at different timepoints relative to GL261-Luc tumor inoculation (d0). Tumor growth kinetics (g-h) and survival (f) was monitored (n = 5 for all groups). *P < 0.05; **P < 0.01; ***P <0.001; ****P<0.0001 (two-sided Log-rank Mantel-Cox test). Data are mean ± S.D. *P < 0.05; **P < 0.01; ***P <0.001; ****P<0.0001 (two-tailed unpaired Student’s t-test)