Fig. 3
CBAP deficiency reduces aerobic glycolysis and energy metabolism and attenuates the Raf-MEK-ERK–RSK and mTORC1-S6K-4E-BP1 signaling pathways. a Functional categories of KEGG gene sets markedly impacted (FDR < 0.25) by CBAP downregulation. b Correlation of CBAP positively-regulated genes and TSC1/TSC2-dependent rapamycin-sensitive genes [26], as determined by GSEA. c qRT-PCR analysis of the mRNA levels of genes involved in the regulation of glycolysis and fatty acid biosynthesis. Gapdh was used as an internal control. The data are averages of biologically triplicated experiments. Results are plotted as mean ± SD (n = 3). d Expression of CBAP, c-Myc, P53 and HIF1α in Jurkat-derived cell clones. e Analysis of concentrations of lactate and α-ketoglutarate in the culture media of Jurkat and CBAP-KO cells. Data are expressed as means ± SD (n = 3; duplicate per measurement). f Suppression of p70S6K phosphorylation in cr-Ctrl and leukemic CBAP-KO Jurkat cells treated with the indicated doses of rapamycin for 6 h. Data represent mean ± SD of three independent experiments with duplicates of each condition. g Effect of the mTORC1 inhibitor, rapamycin, on Jurkat-derived leukemic cell growth. Data represent mean ± SD of three independent experiments. h, i Activation of signaling proteins involved in the Akt-TSC2-mTORC1 (h) and Raf-MEK-ERK (i) signaling pathways in the different Jurkat-derived cell lines, as indicated. Numbers under the lanes represent phosphor-proteins/total proteins normalized to that of cr-Ctrl cells (as 1.0). j, k Effects of the Akt inhibitor MK2206 (j) and the MEK inhibitor U0126 (k) on the growth of cr-Ctrl Jurkat and CBAP-KO leukemic Jurkat cells in tissue culture. Data represent mean ± SD of three independent experiments with duplicates of each condition. *P < 0.05; **P < 0.01; ***P < 0.001, according to two-tailed unpaired Student's t tests
