Fig. 4

HMGB1 is a functional target of miR-142-3p. (a, b) qRT‒PCR analysis of HMGB1 expression in NCI-H3255 cells after transfection with miR-142-3p mimics or a miR-142-3p inhibitor. ***p < 0.001. (c) Western blot analysis of HMGB1 in NCI-H3255 cells after transfection with miR-142-3p mimics or a miR-142-3p inhibitor. (d) Relative luciferase activity of the WT HMGB1 3’ UTR or HMGB1 3’ UTR with mutation of potential binding sites after transfection with miR-142 mimics in 293T cells. *p < 0.05. (e, f) qRT‒PCR to test the expression levels of miR-142-3p and HMGB1 in NCI-H3255 cells transfected with the control, miR-142 mimics, or the HMGB1 overexpression vector and cotransfected with miR-142 mimics and the HMGB1 overexpression vector. **p < 0.01, ***p < 0.001. (g) EdU analysis of the cell proliferation ability of NCI-H3255 cells transfected with the control, miR-142 mimics, the HMGB1 overexpression vector and cotransfected with miR-142 mimics and the HMGB1 overexpression vector. *p < 0.05. The scale bar is 25 μm. (h) Wound healing assays to detect cell migration ability in NCI-H3255 cells transfected with the control, miR-142 mimics, or the HMGB1 overexpression vector and cotransfected with miR-142 mimics and the HMGB1 overexpression vector. *p < 0.05, ***p < 0.001. (i, j) Transwell assay to detect cell migration (i) and invasion (j) of NCI-H3255 cells transfected with the control, miR-142 mimics, or the HMGB1 overexpression vector respectively and cotransfected with miR-142 mimics and the HMGB1 overexpression vector. **p < 0.01, ***p < 0.001. (k) Western blot analysis of HMGB1 and the MAPK/ERK, NF-κB and PI3K/AKT pathways in NCI-H3255 cells transfected with the control, miR-142 mimics, or the HMGB1 overexpression vector and cotransfected with miR-142 mimics and the HMGB1 overexpression vector