Research Paper Volume 16, Issue 11 pp 9846—9858

Figure 3. MIR181A2HG sponged miR-223-3p. (A) Venn diagram illustrates the intersection of upregulated miRNAs from GEO datasets (GSE145054 and GSE142582) and miRDB predicted targets of MIR181A2HG. (B) The binding site between MIR181A2HG and miR-223-3p. HaCaT keratinocytes were co-transfected with miR-223-3p/NC mimic and MIR181A2HG wide type (MIR181A2HG-WT)/MIR181A2HG mutant (MIR181A2HG-MUT) plasmid for 48 hours. Dual-luciferase reporter assay was performed to verify the interaction between MIR181A2HG and miR-223-3p. (C) Expression correlation of MIR181A2HG and miR-223-3p in dataset GSE142582. (D, E) qRT-PCR was used to detect the expression of MIR181A2HG, KRT6 and KRT16 in HaCaT keratinocytes with the transfection of miR-223-3p mimic/inhibitor. GAPDH served as an internal reference. (F) Western blotting was used to detect the protein level of Cyclin A2, CDK4, Cyclin D1, KRT6 and KRT16 in HaCaT keratinocytes with the transfection of miR-223-3p mimic/inhibitor. GAPDH served as an internal reference. (G) The rate of cell proliferation was assessed using EdU incorporation assays. Scale bar: 100 μm. (H) Flow cytometry was performed to analyze the effects on cell cycle distribution. *P<0.05, **P<0.01, ***P<0.001.