Fig. 6: Cav-1 was responsible for increased internalization of HUVECs-derived EVs by neurons treated by OGD.

a Neurons were transfected with three different siRNAs targeting Cav-1. The mRNA level of Cav-1 was determined by qRT-PCR. b The efficiency of si-Cav-1 and si-Cav-3 was further confirmed by Western blotting. c, d Primary neurons were transfected with si-NC, si-Cav-1, or si-Cav-3, and treated with OGD and incubated with HUVECs-derived EVs for 12 h. Cells were observed under a fluorescence microscope, and the mean fluorescence intensity of DiI-labeled EVs in neurons was measured c. miR-1290 was determined in different groups d. e–g Knockdown of Cav-1 abrogated EVs-mediated neuronal protection in OGD, as determined by aCasp-3 Western blotting e, TUNEL f, and LDH release g. h Male mice of 6–8 weeks old were injured by MCAO. Immediately after the MCAO operation, mice were injected intracranially with 1 μL of pKH67-PBS or pKH67-sEVs at the site adjacent to the hippocampus of the impaired hemisphere under the navigation of a murine brain stereotaxic apparatus. The brain samples were sectioned and stained routinely, photographed under a fluorescence microscope, and apoptotic cells of CA1–CA2 were quantitatively compared according to Supplementary Fig. S1. The injection regions that were too far or too close to the hippocampus were excluded from analysis (n = 5 for PBS and n = 6 for EVs). Bars = means ± s.e.m., n = 3. *P < 0.05, **P < 0.01, ***P < 0.001, n.s., not significant.