Fig. 4

cMYC leads to a change in metabolic enzymes in CCA cells. a Heat map of differentially expressed genes (DEGs) in CCA and paired normal samples. The heat map was drawn using the gplots package in Bioconductor. DEGs with Fold Change (FC) > 2 were shown in red; DEGs with Fold Change (FC) < − 2 were in green (P < 0.01) and false discovery rate (FDR) < 0.05. b Gene Ontology (GO) annotation pathways of high and low expression genes in CCA. GO annotation pathways were generated using the ggplot2 package in R language. The size of the dots represents the number of genes. Dot color represents the P-Value. Red: high degree of enrichment, Green: low degree of enrichment. c After cells were transfected with the cMYC plasmid or SiRNA, the concentrations of the metabolites in the cells were measured by Mass spectrometry and normalized to their protein level. d ¹³C₆ glucose labeling experiments were performed as described in the Methods. cMYC-overexpressed and control HuCCT1 cells were treated with ¹³C₆ glucose for 1 h, then relative abundance of ¹³C₃ lactate and ¹³C₄ malate was determined by gas chromatography-mass spectrometer (GC-MS). The incorporation of ¹³C atoms are denoted as m + n, where n is the number of ¹³C atoms. e Cells were transfected with cMYC SiRNA and subjected to western blot. f Cells were transfected with the cMYC plasmid and subjected to western blot. g Cells were transfected with the mutant cMYC plasmid and subjected to western blot. Data represent the Mean ± SEM, n ≥ 3. *p < 0.05, **p < 0.01, NS not significant