Fig. 7

LIN9 binds to the promoter region of AURKA to increase its expression. A, B Western blotting and qRT-PCR experiments were conducted to analyze the effects of IL-1β and PTX on the protein and mRNA expression levels of AURKA and FOXM1 (n = 3). C After knocking down LIN9, western blot experiments to observe whether the promoting effect of IL-1β on downstream molecules was blocked (n = 3). D, E The effects of different drug treatments on the protein expressions of AURKA and FOXM1 were evaluated using western blot assay (n = 3). F Primer design for AURKA was conducted based on the predicted binding sites, and ChIP followed by qRT-PCR experiments were performed to validate the enrichment levels of each binding site (n = 3). G The relative luciferase activity of plasmids containing the wild-type AURKA promoter (WT) or its mutants was measured in 293T cells transfected with plasmids overexpressing LIN9 or control plasmids. The upper sequences show the LIN9 binding motif (blue) and its mutant (red; n = 3). H The differences in the enrichment levels of LIN9 at the AURKA promoter region were observed under different drug treatments (n = 3). Data are presented as mean ± SEM (ns p > 0.05, *p < 0.05, **p < 0.01, ***p < 0.001). I The EMSA was performed to confirm the binding of LIN9 to the A means p (+ 34/+45) and B means p (+ 66/+77) binding sites of AURKA