(A) Immunoblot detection of GLUT1, GLUT3 and MCT4 in the lysates of the Neo or sCPE-transfected LNT229 or Tu140 cells upon CPE knockdown. For Tu140, control siRNA (si-mock) was used as negative control. α-tubulin was used as a loading control. The cells were serum-starved for 24h in serum-reduced medium prior to lysis. A representative immunoblot is shown. (B) Immunohistochemical staining of GLUT1 and LDHA in the Neo or sCPE-overexpressing LNT229 cells (20x magnification, scale bar 100μm). (C) Immunohistochemical staining of GLUT1 in the LNT229 spheres (20x magnification, scale bar 100μm). (D, E) qPCR analysis of CPE, Glut1, Glut3 and MCT4 gene expression in the (D) Neo or sCPE-overexpressing LNT229 cells or (E) Tu140 cells upon CPE knockdown. Control siRNA (si-mock) was used as negative control for CPE knockdown. Red dots represent single experiments. Unpaired t-test with Welch's correction. Mean±SEM; n=3 (D: *p=0.0105; E: *p=0.0422). (F, G) qPCR analysis of the (F) glycolytic enzymes (ALDOC, HKII, PFKP, PFKM, LDHA) and (G) enzymes involved in the pentose-phosphate pathway (G6PD, PGD, TALDO1, TKT, LDHB) in the Neo or sCPE-overexpressing LNT229 cells. Red dots represent single experiments. Unpaired t-test with Welch's correction. Mean±SEM; n=3.