Extended Data Fig. 2: GAPDH(T175A) in MEFs mimics GAPDH(Y314F) in HAP1 cells.
From: The GAPDH redox switch safeguards reductive capacity and enables survival of stressed tumour cells
(a) GAPDH activity as measured in the lysate of MEF cells expressing either WT or mutant (T175A) GAPDH, following treatment of intact cells with 200 µM H2O2 for 5 min. Activity values are normalized relative to untreated WT cells. Based on n = 3 biological replicates. Dotted lines: SD. (b) Hyperoxidation (sulfinylation and/or sulfonylation) of GAPDH Cys-152 in WT and mutant (T175A) MEF cells as visualized by immunoblotting before and after treatment of intact cells with up to 2 mM H2O2 for 5 min. Representative of n = 3 biological replicates. (c) Depletion of extracellular H2O2 (starting concentration: 200 µM) from the cell culture supernatant as measured with an H2O2-selective electrode, in the presence (left and middle panels) or absence (right panel) of 10 mM glucose, and in the presence of 10 mM 6-aminonicotinamide (6AN) (middle panel). Based on n = 3 biological replicates. Dotted lines: SD.
