Nrp-1 Mediated Plasmatic Ago2 Binding miR-21a-3p Internalization: A Novel Mechanism for miR-21a-3p Accumulation in Renal Tubular Epithelial Cells during Sepsis

Figure 6

Interaction of Nrp-1 with either Ago2 or miR-21a-3p. (a) Representative results of immunoprecipitation of the cell lysates of TECs with different treatments; the lysates were immunoprecipitated with Ago2 antibody and probed with Ago2 and Nrp-1 antibodies after electrophoresis. (b) Representative results of RNA pull-down assay for lysates of TECs treated with biotin-miR-21a-3p single strain mimics. Biotin-miR-21a-3p was precipitated; Ago2 and Nrp-1 were probed with immunoblotting. (c) Representative results of RNA pull-down assay for biotin-miR-21a-3p/Nrp-1 mixture. Biotin-miR-21a-3p was precipitated and Nrp-1 was probed with immunoblotting. (d) Representative results of RNA pull-down assay for biotin-miR-21a-3p/Ago2/Nrp-1 mixture. Biotin-miR-21a-3p was precipitated; Ago2 and Nrp-1 were probed with immunoblotting (Ctrl: control; SiNrp-1: Nrp-1 siRNA transfection; SiRNA NC: siRNA negative control transfection; NC: miR-21a-3p mimic negative control; Mimic: miR-21a-3p mimics).

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