Research Paper Volume 13, Issue 6 pp 8127—8145

Long noncoding RNA NKILA transferred by astrocyte-derived extracellular vesicles protects against neuronal injury by upregulating NLRX1 through binding to mir-195 in traumatic brain injury

Astrocytes transfer NKILA into neurons through EVs. (A) NKILA expression in astrocytes detected by RT-qPCR. (B) representative electron micrograph of EVs isolated from astrocytes (scale bar: 100 nm). The red arrow points to the EVs. (C) size and particle distribution plots of isolated EVs from culture medium by NTA. (D) expression of GLT-1, Hsp70, CD63 and Alix as well as the negative marker GRP94 evaluated by Western blot analysis. (E) observation of EVs released by astrocytes under an electron microscope (scale bar: 100 nm). (F) microscopic views of uptake of EVs by neurons (× 400). (G) the expression of NKILA in the supernatant of the astrocyte-derived EVs and EV-free Ast-CM by RT-qPCR; * p p H) uptake of EVs by neurons observed by the confocal laser microscopy (scale bar: 10 μm). All data were measurement data and expressed as mean ± standard deviation. Unpaired t test was used for comparison between two groups. One-way ANOVA, followed by Tukey’s post-hoc test was used for multi-group comparison. The cell experiment was repeated three times.

Figure 6. Astrocytes transfer NKILA into neurons through EVs. (A) NKILA expression in astrocytes detected by RT-qPCR. (B) representative electron micrograph of EVs isolated from astrocytes (scale bar: 100 nm). The red arrow points to the EVs. (C) size and particle distribution plots of isolated EVs from culture medium by NTA. (D) expression of GLT-1, Hsp70, CD63 and Alix as well as the negative marker GRP94 evaluated by Western blot analysis. (E) observation of EVs released by astrocytes under an electron microscope (scale bar: 100 nm). (F) microscopic views of uptake of EVs by neurons (× 400). (G) the expression of NKILA in the supernatant of the astrocyte-derived EVs and EV-free Ast-CM by RT-qPCR; * p < 0.05 compared with the Vector-EVs. # p < 0.05 compared with the treatment of NKILA-EVs. (H) uptake of EVs by neurons observed by the confocal laser microscopy (scale bar: 10 μm). All data were measurement data and expressed as mean ± standard deviation. Unpaired t test was used for comparison between two groups. One-way ANOVA, followed by Tukey’s post-hoc test was used for multi-group comparison. The cell experiment was repeated three times.