Figure 3.
Transferred ILC2s promote angiogenesis and vascular development after ischemic stroke. (A) Experimental design of scRNA-seq of ischemic hemisphere after MCAO and ILC2s transfer (created with https://BioRender.com). (B) UMAP plot of scRNA-seq clustering at 14 days poststroke in the ischemic hemisphere with and without i.v. transfer of ILC2s. MM, monocyte and macrophage; MG, microglia; EC, endothelial cell; OL, oligodendrocyte; AS, astrocytes; DC, dendritic cell; PC, pericyte; CPC, choroid plexus cells; T, T cells; OPC, oligodendrocyte precursor cell; NeuN, neuron; Pro.Cell, proliferating cell; EPEN, ependymal cells; SMC, smooth muscle cell; FB, fibroblast; OEG, olfactory ensheathing glia. (C) Number of DEGs in different cell types (left column), ratio of cell proportion (middle column), and AUCell score for healing (right column), ILC2-transferred group versus PBS group. (D and E) Lectin and Evans blue staining results and the quantification at 14 days after stroke in PBS group (n = 4) and ILC2-transfer group (n = 3). White dashed lines indicate basal ganglion area. The bar plot was analyzed by two-sided, unpaired Student’s t test. All data represent biological replicates from two independent experiments. *P = 0.0334. Scale bar, 500 µm. (F and G) Immunofluorescence of ZO-1 and CD31 staining and ZO-1 coverage quantification at day 14 after stroke (n = 3, 12 views for each group, each dot stands for a view). The bar plot was analyzed by two-sided, unpaired Student’s t test. All data represent biological replicates from two independent experiments. *P = 0.0449. Scale bar, 100 µm. (H and I) Immunofluorescence of lectin-488 staining and quantification for vessel coverage in PBS group (n = 4) and ILC2-transfer group (n = 3) within the basal ganglia region at day 14 after MCAO. The bar plot was analyzed by two-sided, unpaired Student’s t test. All data represent biological replicates from two independent experiments. *P = 0.0166. Scale bar, 200 µm. (J and K) Immunofluorescence of BrdU and CD31 and quantification of BrdU+CD31+ cell proportion in the basal ganglia region the staining results at 14 days after stroke in the two groups (n = 3, 10 views for each group, each dot stands for a view). The bar plot was analyzed by two-sided, unpaired Student’s t test. All data represent biological replicates from two independent experiments. ****P < 0.0001. Scale bar, 100 µm. ZO-1, tight junction-associated proteins.

Transferred ILC2s promote angiogenesis and vascular development after ischemic stroke. (A) Experimental design of scRNA-seq of ischemic hemisphere after MCAO and ILC2s transfer (created with https://BioRender.com). (B) UMAP plot of scRNA-seq clustering at 14 days poststroke in the ischemic hemisphere with and without i.v. transfer of ILC2s. MM, monocyte and macrophage; MG, microglia; EC, endothelial cell; OL, oligodendrocyte; AS, astrocytes; DC, dendritic cell; PC, pericyte; CPC, choroid plexus cells; T, T cells; OPC, oligodendrocyte precursor cell; NeuN, neuron; Pro.Cell, proliferating cell; EPEN, ependymal cells; SMC, smooth muscle cell; FB, fibroblast; OEG, olfactory ensheathing glia. (C) Number of DEGs in different cell types (left column), ratio of cell proportion (middle column), and AUCell score for healing (right column), ILC2-transferred group versus PBS group. (D and E) Lectin and Evans blue staining results and the quantification at 14 days after stroke in PBS group (n = 4) and ILC2-transfer group (n = 3). White dashed lines indicate basal ganglion area. The bar plot was analyzed by two-sided, unpaired Student’s t test. All data represent biological replicates from two independent experiments. *P = 0.0334. Scale bar, 500 µm. (F and G) Immunofluorescence of ZO-1 and CD31 staining and ZO-1 coverage quantification at day 14 after stroke (n = 3, 12 views for each group, each dot stands for a view). The bar plot was analyzed by two-sided, unpaired Student’s t test. All data represent biological replicates from two independent experiments. *P = 0.0449. Scale bar, 100 µm. (H and I) Immunofluorescence of lectin-488 staining and quantification for vessel coverage in PBS group (n = 4) and ILC2-transfer group (n = 3) within the basal ganglia region at day 14 after MCAO. The bar plot was analyzed by two-sided, unpaired Student’s t test. All data represent biological replicates from two independent experiments. *P = 0.0166. Scale bar, 200 µm. (J and K) Immunofluorescence of BrdU and CD31 and quantification of BrdU+CD31+ cell proportion in the basal ganglia region the staining results at 14 days after stroke in the two groups (n = 3, 10 views for each group, each dot stands for a view). The bar plot was analyzed by two-sided, unpaired Student’s t test. All data represent biological replicates from two independent experiments. ****P < 0.0001. Scale bar, 100 µm. ZO-1, tight junction-associated proteins.

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