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

Figure 4. The high expression of miR-4505 in Exos-shVDR promoted macrophage proliferation and M1 polarization while inhibiting macrophage apoptosis.

Figure 4

(A) A volcano plot showing the miRNA expression profile between the psoriatic skin tissues and the normal skin tissues in the GSE115293 expression matrix and bar graphs showing the expression levels of 12 abnormally upregulated miRNAs in normal skin tissue and psoriatic skin tissue. Red circles represent significantly up-regulated miRNAs, blue circles represent significantly down-regulated miRNAs, and black circles represent no differentially expressed miRNAs. N = 3. (B) RT-qPCR was performed to evaluated the differential expression of miR-4507, miR-4653, and miR-4505 between Exos-shVDR and Exos-shNC. N = 3. (C) The expression of miR-4505 in macrophages treated with Exos-shVDR or Exos-shNC was detected by RT-qPCR. N = 3. (D) The expression of miR-4505 in macrophages transfected with miR-4505 mimics or NC mimics was assessed by RT-qPCR. N = 3. (E) Macrophage proliferation ability was detected by the CCK-8 assay. N = 5. (F) Apoptosis was assessed by flow cytometry with AV-FITC/PI staining. N = 3. (G) The expression levels of M1 macrophage markers (IL-1β, TNFα, and IL-6) and M2 macrophage markers (IL-10) in macrophage were assessed by RT-qPCR. N = 3. (H) ELISA was performed to evaluate the content of M1 macrophage markers (IL-1β, TNFα, and IL-6) and M2 macrophage markers (IL-10) in the supernatant of macrophages. N = 5. n.s.P > 0.05, *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-shNC, exosomes derived from HaCaT cells with stably transfected shNC; Exos-shVDR, exosomes derived from HaCaT cells with stably transfected shVDR; NC mim., negative control mimics; miR-4505 mim., miR-4505 mimics.