Schistosoma japonicum extracellular vesicle miRNA cargo regulates host macrophage functions facilitating parasitism (original) (raw)
Fig 3
S. japonicum EV miRNAs are taken up by recipient cells and incorporated into host Argonaute.
(A) Uptake of SjEV RNAs by RAW264.7 cells detected using fluorescence microscopy. SjEV RNAs were labeled using the Exo-Glow exosome labeling kit then incubated with RAW264.7 cells. EVs isolated from a murine liver cell line (NCTC clone 1469 cells) were used as a positive control. Red indicates the labeled SjEV RNAs. Nuclei were stained with 4', 6-Diamidino-2-Phenylindole (blue). Bar indicates 50 μm. (B) Detection of SjEV miRNAs in the recipient cells treated with SjEVs by RT-qPCR. (C) Immunoblotting analysis of Argonaute pull down products using Anti-AGO2 and anti-tubulin antibodies. (D) RT-qPCR identification of SjEV miR-125b and bantam miRNAs are enriched in Argonaute pull down products. For panel B and D, data illustrate representative results and show the mean and standard errors from an experiment carried out in triplicate. * P ≤ 0.05 and ** P ≤ 0.01.