Fig. 7: Effect of the transport protein HNRNPA1 on the exosomal sorting of miR-483-5p in renal TECs. | Cell Death & Disease

Fig. 7: Effect of the transport protein HNRNPA1 on the exosomal sorting of miR-483-5p in renal TECs.

From: HNRNPA1-mediated exosomal sorting of miR-483-5p out of renal tubular epithelial cells promotes the progression of diabetic nephropathy-induced renal interstitial fibrosis

Fig. 7

A The primary mouse renal TECs were cultured under NG and HG conditions for 48 h, and their exosomes were isolated for verification by TEM (Scale Bar = 100 nm) and B western blot (CD63 and CD9 proteins). C Detection of the miR-483-5p levels in cells and exosomes by qRT-PCR. D After HG treatment for 48 h, the RNA pull-down of miR-483-5p was carried out, and the eluate was subjected to mass spectrometry. E Gene Ontology (GO) analysis. F RNA immunoprecipitation verification of HNRNPA1 and HNRNPA2/B1 that bound to miR-483-5p. G RNA pull-down experiment corroborated the binding of HNRNPA1 to miR-483-5p. H After transfecting si-hnrnpa1 into primary mouse renal TECs, the cells were treated with 30 mM HG for 48 h. Detection of the miR-483-5p levels in cells and exosomes by qRT-PCR. I After transfecting si-hnrnpa1 and/or miR-483-5p inhibitor into primary mouse renal TECs, the cells were treated with 30 mM HG for 48 h. Detection of HNRNPA1, CoL I, CoL III, TIMP2, ERK1/2, and pERK1/2 protein levels by western blot. **P < 0.01 vs. NG, IgG or HG + si-NC. ##P < 0.01 vs. Anti-HNRNPA1. Student’s t-test and one-way ANOVA followed by Tukey’s post-test. of three independent experiments. NG normal glucose, HG high glucose, NC negative control.

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