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. 2023 Mar 15;10(14):2206139. doi: 10.1002/advs.202206139

Figure 4.

Figure 4

SOX17 induces the expression of miR‐224‐5p and miR‐361‐3p at the transcriptional level and stimulates the release of miR‐224‐5p and miR‐361‐3p into intercellular space through exosomes. a,b) Heat map and scatter diagram of SOX17‐induced changes in exosome‐associated miRNA expression. Exosomes were collected from HPAECs treated with Ad‐Empty or Ad‐SOX17 to perform unsupervised hierarchical clustering. Each row represents one miRNA, and each column represents one sample. c) qRT‐PCR assay for the expression of the selected six differential miRNAs in SOX17‐associated exosomes, *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001 versus Ad‐Empty exo, n = 4. d) qRT‐PCR assay for the expression of miR‐224‐5p and miR‐361‐3p in HPAECs treated with SOX17‐associated exosomes, **P < 0.01, ***P < 0.001 versus Ad‐Empty exosomes, n = 4. e) RNA‐ISH assay for the expression of miRNA‐224‐5p and miR‐361‐3p in the lung tissues of mice treated with SOX17‐associated exosomes through intravenous injection, values are mean fold‐changes of controls, ***P < 0.001, ****P < 0.0001 versus Ad‐Empty exo, n = 6. f) The DNA motif for SOX17 was obtained from JASPAR. g) The binding sites in the promoter of the host gene of miR‐224‐5p (GABRE) and miR‐361‐3p (CHM) were predicted by miRIAD and JASPAR. (h) The regulation of SOX17 in the promoter of the host gene of miR‐224‐5p (GABRE) and miR‐361‐3p (CHM) were detected by luciferase reporter assay, ***P < 0.001 versus Ad‐Empty, n = 4. i,j) The combination between SOX17 and the promoters of the host gene of miR‐224‐5p and miR‐361‐3p was verified by ChIP and DNA pull‐down assays, ****P < 0.0001 versus Anti‐lgG, n = 3. k) The expression of miR‐224‐5p and miR‐361‐3p in SOX17‐associated exosomes as well as SOX17‐overexpressing HPAECs were examined by qRT‐PCR, ***P < 0.001, ****P < 0.0001 versus Ad‐Empty exo group, ##P < 0.01 versus Ad‐Empty HPA group, n = 4. l) Export ratios of miR‐224‐5p and miR‐361‐3p in SOX17‐associated exosomes relative to SOX17‐overexpressing HPAECs.