G6pd assay kit

The levels of G6PDH- activity were performed according to the manufacturer’s protocols for G6PD assay kit (ab102529, Abcam, Cambridge, MA, USA.]. Sorbitol synchronized trophozoite stage parasites (haematocrit: 2.5%, parasitaemia: 5%) under normal culture conditions were washed with PBS twice and 10⁸ RBC were seeded into a poly-L-lysine-coated 96 well plate. Samples and their controls, and the standard curve for NADPH were read at 450 nm on the FlexStation3.

Following transfection, colon cancer cells (1 × 10⁶ cells/well) were incubated for 24 h. Glucose and lactate levels in the supernatants of cultured cells and its intracellular levels were measured using the Glucose Colorimetric Assay Kit (BioVision, Milpitas, USA) and Lactate Assay kit (BioVision, Milpitas, USA), respectively. Some cells were lyzed and the levels of G6PD enzymatic activity and intracellular NADPH in cell lysates were determined using G6PD Assay Kit (ab102529, Abcam, UK) and Amplite TM Colorimetric NADP/NADPH Ratio Assay Kit (ab65349, Abcam, UK), respectively.

Following transfection, the CRC cells (1×10⁶ cells/well) were incubated for 24 h. Glucose levels in the supernatants of cultured cells and cell lysates were measured using the Glucose Colorimetric Assay Kit (BioVision, Milpitas, USA) and the levels of lactate in the supernatants of cultured cells were quantified by an automatic analyzer. The levels of intracellular G6P in cell lysates were analyzed using a G6P Assay kit (BioVision, Milpitas, USA). In addition, the levels of intracellular G6PD enzymatic activity and NADPH in cell lysates were determined using G6PD Assay Kit (ab102529, Abcam, UK) and Amplite TM Colorimetric NADP/NADPH Ratio Assay Kit (ab65349, Abcam, UK), respectively.