Fig. 6

Neuronal EVs contained VEGFA are involved in brain vascular angiogenesis

(A) Confocal images analysis of VEGF-A expression in mouse primary cortical neurons at 5 days in culture.

(B) Western blot identification of the EVs derived VEGF-A in mouse primary cortical neurons.

(C) Confocal images analysis of rescued-intracerebral-hemorrhage phenotype in VEGF-121 microinjected vps28 mutants in 54 hpf Tg (Kdrl: eGFP; Gata1: DsRed) larvae.

(D) Statistical analysis of DAPI leakage in the VEGF-121 injected embryos corresponding to (C).

(E) Confocal images analysis of CtAs branching in VEGF-121 injected embryos at 54 hpf.

(F) Graphical representations of the CtAs numbers in (E).

(G) qPCR analysis of the vegfaa expression level in vps28 mutants and siblings at 30 hpf and 54 hpf.

(H) Western blot analysis of Vegfa protein level in vps28 depleted embryos compared with vps28 siblings in the whole embryos.

(I) Western blot analysis of Vegfa protein level in EVs of the 54 hpf vps28 mutants compared with relative vps28 siblings.

(J) Effects of vegfaa specific deleted exosomes on vps28 mutants CtAs angiogenesis at 54 hpf.

(K) Graphical representations of the CtAs numbers in (J). Data are represented as mean +/− SD. ∗∗p < 0.01.

(L) Vps28 working model. Vps28 is highly expressed in neurons and participates in neurovascular communication by controlling the secretion of neuronal EVs.