Fig. 5

ZFAS1 enhances OLA1 activity and activates glycolysis by binding to the OBG-type domain of OLA1. a Schematic diagram of ZFAS1 directly binding to OLA1 to enhance its ATPase activity and ultimately promote ATP hydrolysis and glycolysis. b Co-localization of ZFAS1 and OLA1 protein detected by the combination of ISH and IF assays in HCT116 and SW620 cells. Scale bar = 5 μm. c MOE verified that ZFAS1-Wild directly binds OLA1 protein and exposes the ATP-binding site of OLA1 protein. d RNA pull-down followed by western blot showed in vitro binding of the ZFAS1-Wild and ZFAS1-Mutant probes with OLA1 protein after ZFAS1 mutation in HEK293T cells. e Rescue experiments determining the ATPase activity of OLA1 after co-transfected with shOLA1 and ZFAS1-Wild/ZFAS1-Mutant vectors in HCT116 cells. f, g The content of lactate or ATP in the cell supernatant after co-transfected with shOLA1 and ZFAS1-Wild/ZFAS1-Mutant vectors in HCT116 cells. h ECAR experiment verifying the effects of OLA1 and ZFAS1-Wild/ZFSA1-Mutant on glycolysis of CRC cells. Data were shown as mean ± s.d. of at least three independent experiments. *P < 0.05; **P < 0.01; *** P < 0.001; **** P < 0.0001 and ns no significance