miR-196a-5p-Rich Extracellular Vesicles from Trophoblasts Induce M1 Polarization of Macrophages in Recurrent Miscarriage

Figure 2

miR-196a is highly expressed in trophoblast-derived EVs and plasma EVs of RM patients. (a) qPCR analysis of the expression levels of miR-196a-5p in the nucleus, cytoplasm, and CM of the HTR-8 cell lines. (b) qPCR analysis of the expression levels of miR-196a-5p in the CM of HTR-8 cells treated with RNase-A (2 mg/ml) alone or in combination with 0.1% Triton X-100. (c) Identification of the EVs derived from HTR-8 cells using transmission electron microscopy (top left), western blotting (bottom left), and NTA (right). (d) qPCR analysis of miR-196a-5p expression levels in the CM, CM with EVs removed by ultracentrifugation, and EVs of HTR-8 cells. (e) qPCR analysis of miR-196a-5p expression in the CM of HTR-8 cells treated with or without GW4869, an inhibitor of EV secretion, and in the EVs derived from the treated and untreated HTR-8 cells. (f) Identification of EVs isolated from the plasma of HC and RM patients through transmission electron microscopy (top) and western blotting (bottom). (g) qPCR analysis comparing the expression levels of miR-196a-5p in the plasma EVs of 30 RM patients and 30 HC individuals (; ; ; ns: not significant). CM: culture medium; NTA: nanoparticle tracking analysis.

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