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. 2023 Aug 4;11:e15798. doi: 10.7717/peerj.15798

Figure 5. Exos-shVDR promoted macrophage proliferation and M1 polarization while inhibiting macrophage apoptosis via transferring miR-4505.

Figure 5

(A) Exos-shVDR was transfected with NC inhibitor and miR-4505 inhibitor using the Exo-Fect Exosome Transfection kit, and RT-qPCR was applied to assess the expression of miR-4505. N = 3. (B) Exos-shVDR-miR-4505 inhibitor and Exos-shVDR-NC inhibitor were applied to treat macrophages, and the expression of miR-4505 in macrophages was detected by RT-qPCR. N = 3. (C) Macrophage proliferation was measured by the CCK-8 assay. N = 5. (D) Macrophages apoptosis was assessed by flow cytometry with AV-FITC/PI staining. N = 3. (E) The expression levels of M1 macrophage markers (IL-1β, TNFα, and IL-6) in macrophages were evaluated by RT-qPCR. N = 5. (F) ELISA was applied to evaluate the content of M1 macrophage markers (IL-1β, TNFα, and IL-6) in the supernatant of macrophages. N = 3. n.s.P > 0.05, **P < 0.01, and ***P < 0.001. NC, negative control; shRNA, short hairpin RNA; VDR, vitamin D receptor; mRNA, messenger RNA; miRNA, microRNA; IL, interleukin; TNF, tumor necrosis factor; RT-qPCR, real-time quantitative polymerase chain reaction; ELISA, enzyme-linked immunosorbent assay; AV-FITC/PI, Annexin V-fluorescein isothiocyante/propidium iodide; Exos-shVDR-NC inhibitor, Exos-shVDR transfected with NC inhibitor; Exos-shVDR-miR-4505 inhibitor, Exos-shVDR transfected with miR-4505 inhibitor.