Article
Extended Data Fig. 4 | VEGF-C signals specif ically in lymphatic endothelial
cells in the meninges and deep cervical lymph nodes and provides survival
benefits in a manner that depends on the administration time point.
a, Gating strategy for LECs and BECs. b, Concatenated FACS plots of LECs and
BECs from meninges and lymph nodes, depicting AKT phosphorylation
intensity. The experiment was repeated independently with similar results.
c, Quantification of the AKT(pS473)-positive population and mean
f luorescence intensity (MFI) within LECs and BECs in the meninges and
deep cervical lymph nodes (meninges: wild type, n = 5; A AV-VEGF-C,
tumour + Luc mRNA, tumour + VEGFC mRNA, n = 8; lymph nodes: wild type,
n = 5; A AV-VEGF-C, n = 8; tumour + Luc mRNA, n = 7; tumour + VEGFC mRNA,
n = 8). d, Fluorescence microscopy images of deep cervical lymph nodes after
treatment with VEGFC mRNA in tumour-bearing mice (CD31, red; LYVE1, green;
DAPI, blue). e, Fluorescence microscopy images of meninges after treatment
with VEGFC mRNA in tumour-bearing mice (CD31, red; LYVE1, green; DAPI,
blue). The experiment was repeated independently with similar results.
f–h, Mice were treated with A AV-VEGF-C or VEGFC mRNA at different time
points relative to GL261-Luc tumour inoculation (day 0). Tumour growth
kinetics (g, h) and survival (f) were monitored (n = 5 for all groups, no tx refers
to mice receiving no treatment). Data are mean ± s.d. *P < 0.05; **P < 0.01;
***P < 0.001; ****P < 0.0001 (two-tailed unpaired Student’s t-test or two-sided
log-rank Mantel–Cox test).