Fig. 4.

Upregulation of miR-744 expression in LPS-treated mast cell-derived exosomes increased miR-744 levels and mitigated ferroptosis in HBE cells. (A) Flowchart of experimental design. LPS-MC-EXs were transfected with either a miR-NC(miR-NC_ LPS-MC-EXs) or miR-744(miR-744_ LPS-MC-EXs), and then incubated with HBE cells. The experiments were performed with three independent technical replicates. (B) qPCR results for miR-744 expression in LPS-MC-EXs after transfection with miR-744. (C–H) Analysis of HBE cells after incubation with LPS-MC-EXs transfected with miR-744. (C) qPCR results showing miR-744 expression. (D) qPCR results for GPX4, ACSL4, and ALOX15 expression. (E) qPCR results for IL6 and TNFa expression. (F) Left panel: Representative western blots for GPX4, ACSL4, and ALOX15; Right panel: Blots were quantified using ImageJ. (G) Flow cytometry data for ROS, including representative images (left) and quantitative analysis (right). (H) Flow cytometry data for apoptosis, including representative images (left) and quantitative analysis (right). ∗P < 0.05, ∗∗P < 0.01, ∗∗∗P < 0.001 (ANOVA, followed by multiple comparison tests). ACSL4, long-chain acyl-CoA synthetase 4; ALOX15, 15-lipoxygenase; GPX4, glutathione peroxidase 4; HBE, human bronchial epithelial; LPS, lipopolysaccharide; miRNA, microRNA; qPCR, quantitative real-time PCR; LPS-MC-EX, exosome from HMC-1 cells stimulated with LPS; WB, Western blot.