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. 2021 May 17;11:666391. doi: 10.3389/fonc.2021.666391

Figure 2.

Figure 2

CircHECTD1 regulated the cell proliferation and migration. (A) RT-qPCR analysis of circHECTD1 expression. (B, C) The expression of proliferation marker genes, CyclinD1, CDK2, and PCNA, were measured by RT-qPCR and western blot. (D) Flow cytometric analysis of the indicated cells transfected with the normal control or circHECTD1. (E) Representative micrographs and quantification of EdU incorporating-cells. Nuclei were stained with Hoechst dye. Bar = 200μm. (F) Representative micrographs and quantification of crystal violet-stained cell colonies. (G) The cells migrating to the lower surface of the membrane were photographed and counted. Bar = 200μm. (H) Photographs were taken when the wound was created at 0 h, 24 h, 48 h, and 72 h in C6 cells, and the percentages of cells migrating into the wound were counted. Bar = 400μm. (I) RT-qPCR analysis depicts the expression of miR-320-5p in the control and circHECTD1 group. (J) Luciferase activity of LUC-circHECTD1 wild-type and mutant in C6 cells transfected with the miR-320-5p mimic and negative control mimic. The data represent the mean ± SEM of three independent experiments. *P < 0.05; ** P < 0.01; *** P < 0.001.