Single nucleus analysis of brain cell transcriptome in adult AAV-CTRL and AAV-VEGF-C mice. (A) Subclustering of forebrain neuronal cells, endothelial cells, vascular mural cells, and immune cells. Top panel: tSNE representation of the neurons after sub-clustering and isolated mapping. tSNE representation of marker gene expression in the neuronal cells subclusters. Middle panel: tSNE representation of vascular mural cells and endothelial cells clusters and distribution of relevant marker genes in tSNE representations. Endothelial cells (Cldn5+), smooth muscle cells (SMCs) (Myh11+); vascular fibroblast (Col1a1+); Pericytes (Pdgfrβ+ Myh11−). Lower panel: tSNE visualization of subclusters of immune cells. Scaled distribution of marker genes of microglia (Tmem119+), monocytes-macrophages (Msr1+, CD68+), and T cells (Cd8b1+). (B) Violin plots representing transcript number in each cluster, between regions (cortex versus striatum, top left). Between conditions (AAV-VEGF-C versus AAV-CTRL, top right) (n = 5 mice/group). Representative histogram of cell numbers in each cluster are shown in the bottom panel. (C and D) Quantification of the levels of BDNF (C) and NGF (D) detected by ELISA in brain sample lysates from 2- to 4-wk-treated mice with AAV-VEGF-C or -CTRL. *P < 0.05, n = 4 mice/group at 2 wk, n = 8–11/group at 4 wk. One-way ANOVA and Tukey’s multiple comparison test. Data are represented as mean ± SEM.
Figure S4.

Single nucleus analysis of brain cell transcriptome in adult AAV-CTRL and AAV-VEGF-C mice. (A) Subclustering of forebrain neuronal cells, endothelial cells, vascular mural cells, and immune cells. Top panel: tSNE representation of the neurons after sub-clustering and isolated mapping. tSNE representation of marker gene expression in the neuronal cells subclusters. Middle panel: tSNE representation of vascular mural cells and endothelial cells clusters and distribution of relevant marker genes in tSNE representations. Endothelial cells (Cldn5+), smooth muscle cells (SMCs) (Myh11+); vascular fibroblast (Col1a1+); Pericytes (Pdgfrβ+ Myh11−). Lower panel: tSNE visualization of subclusters of immune cells. Scaled distribution of marker genes of microglia (Tmem119+), monocytes-macrophages (Msr1+, CD68+), and T cells (Cd8b1+). (B) Violin plots representing transcript number in each cluster, between regions (cortex versus striatum, top left). Between conditions (AAV-VEGF-C versus AAV-CTRL, top right) (n = 5 mice/group). Representative histogram of cell numbers in each cluster are shown in the bottom panel. (C and D) Quantification of the levels of BDNF (C) and NGF (D) detected by ELISA in brain sample lysates from 2- to 4-wk-treated mice with AAV-VEGF-C or -CTRL. *P < 0.05, n = 4 mice/group at 2 wk, n = 8–11/group at 4 wk. One-way ANOVA and Tukey’s multiple comparison test. Data are represented as mean ± SEM.

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