Fig. 6

An EGF-c-Myc-MNX1-AS1 axis promotes the Warburg effect through non-glycolytic functions of PKM2. a-d Huh7 and HCCLM3 cells were infected with pCDH or pCDH-MNX1-AS1 lentiviruses, and HepG2 and PLC/PRF/5 cells were infected with sh-ctrl or sh-MNX1-AS1 lentiviruses. Huh7 and HepG2 cells were subjected to qPCR to determine the expression of LDHA, PDK1 and GLUT1 (A and C). Huh7, HCCLM3, HepG2 and PLC/PRF/5 were subsequently subjected to Western blotting to determine the expression of indicated proteins (B and D). e GSEA analysis of the GLYCOLYSIS pathway in sh-ctrl versus sh-MNX1-AS1 infected HepG2 cells. f MNX1-AS1 levels were measured by qPCR in HepG2, PLC/PRF/5, Huh7 and HCCLM3 cells after treatment with EGF (100 ng/ml) for 24 hours. g and h HepG2 cells infected with sh-ctrl or sh-c-Myc lentiviruses were subjected to mock or EGF (100 ng/ml) treatment for 24 hours before qPCR analysis of RNA level of MNX1-AS1, LDHA, PDK1 and GLUT1 (G) and Western blotting to determine the expression of indicated proteins (H). i and j Huh7 cells infected with sh-ctrl or sh-c-Myc lentiviruses were subjected to infection of pCDH control or pCDH-MNX1-AS1 lentiviruses. qPCR analysis of RNA level of LDHA, PDK1 and GLUT1 (I) and Western blotting to determine the expression of indicated proteins (J). k and l HepG2 cells infected with sh-ctrl or sh-MNX1-AS1 lentiviruses were subjected to transfection of Flag control or Flag-c-Myc. qPCR analysis of RNA level of LDHA, PDK1 and GLUT1 (K) and Western blotting to determine the expression of indicated proteins L. A, C, F, G, I and K are mean ± SD; n = 3 independent experiments, two-way ANOVA with Bonferroni’s multiple comparison post-test, ns, not significant, *p < 0.05, **p < 0.01, ***p < 0.001. B, D, H, J and L are represent of three independent experiments