Fig. 3

MiR-130b can be transferred into AT2 cells co-cultured with BMSCs and increases protein level α/γ-ENaC in AT2 cells. a Transfected miR-130b labelled with Cy3 was localized in AT2 cells co-cultured with BMSCs for 24 h. BMSCs were transiently transfected with Cy3-miR-130b after which they were co-cultured with mouse AT2 cells and then fixed. Nuclei were counterstained by 4,6-diamidino-2-phenylindole (DAPI, blue; left panel), and the representative staining images were shown in CY3-miR-130b positive AT2 cells (red; middle panel). The merged images were seen (right panel). Scale bar, 10 μm. b The result of real-time PCR assays shows miR-130b level in AT2 cells treated with LPS for 12 h and/or co-cultured with BMSCs for 24 h. The relative level of miR-130b were calculated as miR-130b/U6 ratio. c Representative Western blot bands of α- and γ-ENaC protein expression in AT2 cells transfected with miR-130b mimic/inhibitor for 72 h. Blots for β-actin were used as internal controls. d, e Graphical representation of data obtained from Western blot assays for α- and γ-ENaC subunits. Bands were quantified using gray analysis (α-ENaC/β-actin and γ-ENaC/β-actin). **P < 0.01, compared with control; ^&P < 0.05, compared with LPS; ^#P < 0.05, compared with miR-130b mimic negative control (NC); ^$P < 0.05, compared with miR-130b inhibitor negative control (Inhibitor NC), n = 4–6