VEGF-C–producing myeloid cells interact with cpLVs after tMCAO. (A–C) CP sections from sham (A) and tMCAO (B) mice were stained with VEGF-C and Lyve-1 antibodies 7 days after surgery (peak lymphangiogenesis). VEGF-C⁺ cells were observed near Lyve-1⁺ vasculature in sham and tMCAO mice (yellow arrowheads). VEGF-C fluorescence within Lyve-1⁺ cpLVs was measured between groups. Mean fluorescent intensity (MFI) of VEGF-C in Lyve-1⁺ regions were quantified (C) (n = 3 mice per group; mean ± SEM, *P < 0.05, unpaired Student’s t test). Scale bars = 50 µm, 10 µm. (D) Cell-to-cell interactions between cpLECs and immune cells were studied by gating for live doublets from the CP cell suspensions. cpLECs were gated as doublets, live, and CD45⁺, CD31⁺, and PDPN⁺. Macrophages were gated as CD45^hi, CD11b⁺, and CD11c⁻. DCs were gated as CD45^hi, CD11b⁺, and CD11c⁺. CD4 T cells were gated as CD45^hi, CD11b⁻, CD11c⁻, and CD4⁺. CD8 T cells were gated as CD45^hi, CD11b⁻, CD11c⁻, and CD8⁺. B cells were gated as CD45^hi, CD11b⁻, CD11c⁻, CD4⁻, CD8⁻, and B220⁺. cpLECs from singlets were gated as singlets, live, and CD45⁻, CD31⁺, and PDPN⁺. (E) Quantitation of number of cpLECs in singlets, number of cpLECs in doublets, and number of immune cells in doublets near the CP between sham and tMCAO groups after 7 days. Values presented as fold change normalized relative to healthy (n = 3 mice per group; mean ± SEM, *P < 0.05, **P ≤ 0.01, unpaired Student’s t test). (F) Representative orthogonal sectional view of CP after 7 days of tMCAO showing CD11b⁺, CD11c⁺ DCs are bound to Lyve-1⁺ lymphatic vessels. Scale bar = 10 µm. (G) Representative sectional images of CP areas after immunolabeling with CD11b, CD11c, VEGF-C, and Lyve-1 to visualize immune cells and lymphatic vessels after 7 days of sham and tMCAO. Scale bars = 10 µm. BV = blood vessels. Source data are available for this figure: SourceData F4.