Fig. 7: AVIL regulates the stability of FOXM1.

a Whole transcriptome analyses of U87 cells transfected with siAVIL vs. siCT, and stable AVIL overexpression cells vs. empty vector control (CT). b Gene Set Enrichment Analysis revealed highly similar profiles between FOXM1 targets, and differentially expressed genes caused by AVIL silencing and overexpression. NES score = 2.5. c BART prediction ranked FOXM1 as the top factor. d FOXM1 expression can rescue at least some of the AVIL effect. U87 cells transfected with siAVIL or siCT were further transfected with FOXM1 expression vector, or control plasmid (CT). Cell proliferation was measured by cell counting. Cell motility was measured by wound-healing assay. e FOXM1 RNA expression was not affected by AVIL silencing or overexpression. qRT-PCR measuring FOXM1 mRNA level in U87 and U251 cells. FOXM1 RNA expression was normalized against that of GAPDH. No statistical significance (ns) was observed in all groups compared to CT or siCT. f FOXM1 protein was reduced upon AVIL silencing, and induced upon AVIL overexpression. Western blot measured the protein levels of AVIL, FOXM1, and GAPDH. Upper band in AVIL Western is GFP-tagged AVIL. g FOXM1 protein half-life was reduced upon AVIL silencing, and induced upon AVIL overexpression. U87 cells were treated with cycloheximide to inhibit new protein synthesis. FOXM1, and GAPDH proteins were measured by Western Blot analysis. Protein signals were quantified by densitometry. h The reduced half-life of FOXM1 can be offset by the proteasome inhibitor, MG132. Data are presented as mean values ± SD in d, and e.