tMCAO induces lymphangiogenesis at the CP lymphatic vasculature. (A) Experimental timeline. Sham and 60-min tMCAO surgeries were performed on 10–12-wk-old male mice (25–29 g). Across three time points (days 3, 7, and 14), T2-weighted MRIs of the infarct area and multiple tissue regions were collected for lymphatic analysis of the CP, optic nerve, NP, dura, and lymph nodes (created using https://BioRender.com). (B) Representative images to demonstrate the position of Lyve-1⁺ cpLVs which are positioned between OB from a sham and tMCAO mouse on day 7 after surgery. Sections were stained with CD11b (red) to visualize myeloid cells and Lyve-1 (white) to visualize cpLVs. Dotted boxes outline Lyve-1⁺ cpLVs. Scale bars = 300 µm. (C) T2-weighted MRI images of brain sections between sham and tMCAO at days 3, 7, and 14. Representative images of MRI brain scans after sham and tMCAO. White dashed lines mark brain infarcts, which are quantified into a graph (n = 8 mice for sham, n = 9 mice for tMCAO; mean ± SEM, **P ≤ 0.01, ****P ≤ 0.0001, two-way ANOVA). (D) 60-µm coronal sections of CP areas were immunolabeled for Lyve-1 to visualize lymphatic vessels after 3, 7, and 14 days of sham or tMCAO and quantified. Representative confocal images of lymphatic vessels near the CP. Quantitation of average Lyve-1⁺ vessel area near the CP (n = 8 mice for sham, n = 9 mice for tMCAO; mean ± SEM, ***P ≤ 0.001, ****P ≤0.0001, two-way ANOVA). Scale bars = 100 µm. (E) Quantitation of number of lymphatic loops, sprouts (see representative image, day 3 tMCAO) and length near the CP after sham and tMCAO at days 3, 7, and 14 (n = 8 mice for sham, n = 9 mice for tMCAO; mean ± SEM, *P ≤ 0.05, **P ≤ 0.01, ***P ≤ 0.001, ****P ≤ 0.0001, two-way ANOVA). Scale bars = 20 µm. Source data are available for this figure: SourceData F1.