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. 2021 Nov 9;16(6):1347–1364. doi: 10.1002/1878-0261.13125

Fig. 5.

Fig. 5

miR‐485‐5p has a positive effect on senescence and apoptosis in both A549 and H1299. (A) qRT‐PCR to test the expression of miR‐485‐5p in A549 cells transfected with miR‐485‐5p mimic and the NC. U6 serves as the internal control. (B) CCK‐8 assay in miR‐485‐5p‐overexpressing (OE) A549 cells. (C) SA‐β‐Gal staining in miR‐485‐5p‐ OE A549 cells. Images were captured at 100× magnification under a microscope (Left) and the percentage of staining positive cells was quantified (Right). Scale bars = 100 μm. *** represents P < 0.001 based on t‐test with three independent countings. (D, E) Cell cycle analysis showing flow cytometric distribution (D) and quantitative statistics (E) for cells of each cell phase. Cells were synchronized by 2.5 μm colchicine before transfection and cell cycle measurement. (F, G) Apoptosis analysis with double‐staining Annexin V and PI in miR‐485‐5p‐OE A549 cells by flow cytometer (F). Annexin V‐positive cells were considered as apoptotic cells (G). (H) qRT‐PCR to validate the increased expression of miR‐485‐5p after transfecting its mimic into H1299 cells. U6 serves as the internal control. (I) CCK‐8 assay in miR‐485‐5p‐OE H1299 cells. (J) SA‐β‐Gal staining in miR‐485‐5p‐OE H1299 cells. Images were captured at 100× magnification under a microscope (Left) and the percentage of positive staining cells was quantified (Right). Scale bars = 100 μm. ** represents P < 0.01 based on t‐test with three independent countings. (K, L) Cell cycle analysis shown by flow cytometric distribution (K) and quantitative statistics (L) for cells of each phase. Cells were synchronized by 2.5 μm colchicine before transfection and cell cycle measurement. (M, N) Apoptosis analysis by double‐staining Annexin V and PI in miR‐485‐5p‐OE H1299 cells by flow cytometer (M). The proportion of apoptotic cells is indicated by the percentage Annexin V‐positive cells (N). For each panel, *, **, *** represent P < 0.05, P < 0.01, and P < 0.001, respectively, based on t‐test with three biological replicates. Error bars indicated mean ± SEM.