Downregulation of an Evolutionary Young miR-1290 in an iPSC-Derived Neural Stem Cell Model of Autism Spectrum Disorder

Figure 4

mir-1290 is secreted in extracellular vesicles (EVs). (a) Top: transmission electron microscopy (TEM) of EVs isolated from NSCs shows the presence of different sizes of EVs isolated from control-NSC culture media. Bottom: western blot of EVs from NSC whole cell lysate (lysate) and undifferentiated (undiff) and differentiated (diff) EVs shows the presence of EV markers Alix, Flotillin, and Tsg101 enriched in the EV fraction. (b, c) Nanoparticle tracking analysis (NTA, nanosight) revealed no significant differences in the concentration of the particles between control- and ASD-NSC-derived EVs ( donors). (d) RT-PCR data indicates that miR-1290 levels are significantly upregulated in EVs isolated from day 21-differentiated ASD-NSC when compared to the EVs isolated from control-NSCs. # represents significance. (days 7 and 14) and (day 21) ( donors), determined by an unpaired -test and corrected by the Holm-Sidak multiple correction post hoc test. (e) Undifferentiated ASD-NSCs were plated and differentiated for 21 days in the presence of miR-1290, miR-124, and negative mimics complexed with DOTAP followed by immunostaining with neuronal marker MAP2. Results indicate a significant increase in MAP2-expressing cells only in ASD cultures treated with DOTAP-miR-1290. Bar = 20 μm. ( donors), determined by two-way ANOVA followed by Sidak’s multiple correction test. Data are represented as the .

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