Fig. 3

CircHERC1 directly binds miR-142 and regulates miR-142 expression in NSCLC cells. (a) qRT‒PCR for the expression of circHERC1 in NCI-H3255 cells after the pulldown of Ago2 by RIP assay. **p < 0.01. (b) qRT‒PCR of the candidate miRNA expression in circHERC1-overexpressing NCI-H3255 cell lysates pulled down and enriched with a biotinylated circHERC1-specific probe. *p < 0.05, **p < 0.01. (c) The relative luciferase activity of WT circHERC1 3^’ UTR or mutant circHERC1 3’ UTR after transfection with miR-142 mimics in 293T cells. **p < 0.01. (d, e) qRT‒PCR was used to test the expression of circHERC1 and miR-142-3p in NCI-H3255 cells transfected with circHERC1 overexpression vector and miR-142 mimics. **p < 0.01, ***p < 0.001. (f) EdU analysis of the cell proliferation ability of NCI-H3255 cells transfected with the control, the circHERC1 overexpression vector, or miR-142 mimics and cotransfected with the circHERC1 overexpression vector and miR-142 mimics. *p < 0.05. The scale bar is 25 μm. (g) Wound healing assay to detect cell migration ability in NCI-H3255 cells transfected with the control, the circHERC1 overexpression vector, or miR-142 mimics and cotransfected with the circHERC1 overexpression vector and miR-142 mimics. **p < 0.01. (h, i) Transwell assay to detect cell migration (h) and invasion (i) of NCI-H3255 cells transfected with the control, the circHERC1 overexpression vector, or miR-142 mimics and cotransfected with the circHERC1 overexpression vector and miR-142 mimics. **p < 0.01, ***p < 0.001. (j) Western blot analysis of the MAPK/ERK, NF-κB and PI3K/AKT pathways in NCI-H3255 cells transfected with the control, the circHERC1 overexpression vector, or miR-142 mimics and cotransfected with the circHERC1 overexpression vector and miR-142 mimics