Fig. 7

Inhibition of HMGB1 and FOXO1 expression attenuates the influence of circHERC1 on cell proliferation, apoptosis, migration and invasion. (a) Western blot analysis of HMGB1, FOXO1 and the MAPK/ERK, NF-κB and PI3K/AKT pathways in NCI-H3255 cells transfected with the control vector, or the circHERC1 overexpression vector and cotransfected with the circHERC1 overexpression vector and the HMGB1 knockdown vector; the circHERC1 overexpression vector and the FOXO1 knockdown vector; or the circHERC1 overexpression vector, the HMGB1 knockdown vector, and the FOXO1 knockdown vector. (b) EdU analysis of the cell proliferation ability in NCI-H3255 cells transfected with the control vector, or the circHERC1 overexpression vector and cotransfected with the circHERC1 overexpression vector and the HMGB1 knockdown vector; the circHERC1 overexpression vector and the FOXO1 knockdown vector; or the circHERC1 overexpression vector, the HMGB1 knockdown vector, and the FOXO1 knockdown vector. *p < 0.05. The scale bar is 25 μm. (c) Flow cytometry to detect apoptosis of NCI-H3255 cells transfected with the control vector, or the circHERC1 overexpression vector and cotransfected with the circHERC1 overexpression vector and the HMGB1 knockdown vector; the circHERC1 overexpression vector and the FOXO1 knockdown vector; or the circHERC1 overexpression vector, the HMGB1 knockdown vector, and the FOXO1 knockdown vector. *p < 0.05, ***p < 0.001. (d, e) Transwell assay to detect cell migration (d) and invasion (e) of NCI-H3255 cells transfected with the control vector, or the circHERC1 overexpression vector and cotransfected with the circHERC1 overexpression vector and the HMGB1 knockdown vector; the circHERC1 overexpression vector and the FOXO1 knockdown vector; or the circHERC1 overexpression vector, the HMGB1 knockdown vector, and the FOXO1 knockdown vector. *p < 0.05, **p < 0.01, ***p < 0.001