GAPDH and cancer: It was concluded that (i) inhibition of GAPDH with iodoacetate, or PYK/ENO with oxamate but not LDH, PGK, or PGAM, induces Fru1,6BP and DHAP accumulation and methylglyoxal production, leading to significant suppression of glycolysis; and (ii) uncompetitive inhibition of the most controlling pathway steps is the most direct and potent mechanism to efficiently perturb cancer glycolysis.