Fig. 3

MNX1-AS1 promotes glycolysis in HCC. a and b MNX1-AS1 levels were measured by qPCR after infection of Huh7 cells with either pCDH control or pCDH-MNX1-AS1 lentiviruses (A), or after infection of HepG2 cells with lentiviruses containing control shRNA (sh-ctrl) or two independent shRNAs targeting MNX1-AS1 (B). c and d Huh7 (C) and HepG2 (D) cells from (A) and (B) were seeded in 12 wells and cultured in DMEM for 12 hr. Acidification of the culture medium was evaluated by visual inspection. e and f Extracellular lactate production in Huh7 (E) and HepG2 (F) cells from (A) and (B). (g and h) Glucose uptake were measured in Huh7 (G) and HepG2 (H) cells from (A) and (B). i-l ECAR measurements using Seahorse XF assays in Huh7 (I and J) and HepG2 (K and L) cells from (A) and (B). The glycolysis rate were calculated as: (Maximum ECAR before Oligomycin injection)-(Last ECAR before Glucose injection) (J) and (L). A, E, G and J are mean ± SD; n = 3 independent experiments, two-tailed paired Student’s t test, **p < 0.01. B, F, H and L are mean ± SD; n = 3 independent experiments, one-way ANOVA with Tukey’s multiple comparison post-test, **p < 0.01, ***p < 0.001. C and D are represent of three independent experiments. I and K are mean ± SD; n = 3 independent experiments, two-way ANOVA with Bonferroni’s multiple comparison post-test, ns, not significant, **p < 0.01, ***p < 0.001