Fig. 2.

Screening and validation of miR-744 as a key regulator of ferroptosis in mast cell-derived exosomes. (A) Flowchart of the experimental procedure. HMC-1 cells were stimulated with LPS or PBS, and exosomes were collected for miRNA sequencing. Data were analyzed using R (4.4.1). (B) Exosomal miRNA sequencing revealed differential expression of miRNA in PBS-MC-EXs and LPS-MC-EXs. (C) Volcano plot of differentially expressed miRNAs. (D) Heatmap of the top 54 miRNAs (P < 0.05) in PBS-MC-EXs and LPS-MC-EXs. (E) Intersection analysis of top 54 miRNAs along with miRNAs regulating the ACSL4/ALOX15/GPX4 axis. (F) Ferroptosis-associated genes whose expression is regulated by miR-744. (G) Target genes of miR-744 are highlighted with ferroptosis-associated genes in red. (H) Dual-luciferase assay results. ACSL4, long-chain acyl-CoA synthetase 4; ALOX15, 15-lipoxygenase; ARDS, acute respiratory distress syndrome; GPX4, glutathione peroxidase 4; HBE, human bronchial epithelial; LPS, lipopolysaccharide; LPS-MC-EX, exosome from HMC-1 cells stimulated with LPS; PBS-MC-EX, exosome from HMC-1 cells stimulated with PBS. (For interpretation of the references to colour in this figure legend, the reader is referred to the Web version of this article.)