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. 2022 Aug 16;2022:6329097. doi: 10.1155/2022/6329097

Figure 5.

Figure 5

circCNN2 upregulates E2F1 expression via miR-184. (a) RT-qPCR was used to examine the expression of 8 candidate mRNAs (H2AFX, LSM4, ARHGAP39, CLUH, MDH2, E2F1, CDCA4, and TIMM13) upon circCNN2 knockdown. (b, c) RT-qPCR and western blot assays were conducted to test the expression and protein levels of E2F1 upon circCNN2 knockdown in H1703 and H226 cells. (d) RT-qPCR and western blot analysis were used to examine the expression and protein levels of E2F1 in LUSC cells. (e) RT-qPCR and western blot assays were conducted to detect the expression and protein levels of E2F1 upon miR-184 downregulation. (f) RNA pull down assay was carried out to verify the binding ability between miR-184 and E2F1 with Bio-NC as the control. (g) The binding ability between miR-184 and E2F1 was verified through luciferase report assay by testing the luciferase activity of LUSC cells transfected with wild-type or mutated E2F1 plasmids upon mimics NC and miR-184 mimics. (h) Rescue assays were conducted to verify the interaction among circCNN2, miR-184 and E2F1. ∗∗P < 0.01 compared to the control group.