Fig. 1. Downregulation of ASPP2 enhances mevalonate pathway gene expression and cholesterol biosynthesis in HCC cells.

a Top: KEGG Pathway analysis of ASPP2 knockdown show significant differential changes (P < 0.05) in canonical metabolic pathways. The P value for each pathway is indicated by the bar and is expressed as −1 times the log of the P value. Arrows are used to indicated the two pathways closely related with mevalonate metabolism. Bottom: Gene Ontology (GO) analysis revealed genes were most significantly upregulated and enriched by ASPP2 knockdown in the terpenoid backbone biosynthesis pathway (P < 0.05, FDR < 0.25). b Schematic of the mevalonate pathway with asterisk (*) marking upregulated genes by ASPP2 knockdown in panel (a) bottom. c qRT-PCR of Hmgcs1, Hmgcr mRNA expression in HCC-LM3 and HepG2 cells infected with lentivirus encoding shRNA was indicated for 72 h. β-actin was used as a control. d Corresponding western-blot of HMGCS1 and HMGCR in HCC-LM3 and HepG2 cells. e Total cholesterol and free cholesterol were measured by GC-MS methods, using the internal standard strategy as methods described. Each symbol represents the mean ± SD of three wells. Asterisk (*) indicates P < 0.05