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The activity of endogenous phosphofructokinase was measured using Phosphofructokinase Activity Colorimetric Assay Kit (Bio Vision, catalog K776). In brief, 2 × 106 cells were lysed in 100 μl of PFK Assay Buffer on ice. Small molecules in samples were removed by 10 kD spin column (Abcam, catalog ab93349) and 5 μl of lysate was used per assay. Absorbance was measured at 450 nm for two time points. The activity of PFK-1 was calculated by dividing the amount of generated NADH by the reaction time and was normalized against protein concentration.
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