Fig. 4: MiR-182-5p transmitted by MSC-exosomes reverses EMT process by directly targeting Ikbkb.

a Online starBase v2.0 predicted 71 miRNAs targeted to Ikbkb were subjected to RT-qPCR analysis in LPS-treated MLE-12 cells under MSC coculture or not. b RT-qPCR analyzed the level of miR-182-5p in LPS-treated MLE-12 cells treated with MSC-exosome. c Relative level of miR-182-5p was detected by RT-qPCR in LPS-treated MLE-12 cells transfected with specific miRNA mimics. d Flow cytometry analysis displayed the apoptosis rate in LPS-treated MLE-12 cells transfected with miR-182-5p mimics. e IF assay analyzed the fluorescence intensities of two EMT-related proteins, E-cadherin, and Vimentin, in LPS-treated MLE-12 cells transfected with miR-182-5p mimics. Scale bar = 50 μm. f, g RT-qPCR and western blot examined the levels of epithelial marker (E-cadherin) and mesenchymal markers (α-SMA, TGF-β1, Collagen type I, and Collagen type III) in LPS-treated MLE-12 cells transfected with miR-182-5p mimics. h The binding sites between miR-182-5p and Ikbkb were predicted. i Luciferase reporter assay examined the luciferase activity of indicated reporter vectors in LPS-treated MLE-12 cells and HEK-293T cells co-transfected with miR-182-5p mimics or NC mimics. j IF assay detected p65 nuclear translocation in LPS-treated MLE-12 cells with MSC-exosome or MSC-exosome+miR-182-5p inhibitor. Scale bar = 50 μm. k Western blot analysis detected cytoplasmic and nuclear p65 in LPS-treated MLE-12 cells with MSC-exosome or MSC-exosome+miR-182-5p inhibitor. l RT-qPCR and western blot examined the mRNA and protein levels of Ikbkb in LPS-treated MLE-12 cells transfected with MSC-exosome or MSC-exosome+miR-182-5p inhibitor. ^**p < 0.01. n.s. no statistical significance.