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Description

The resulting metabolic stress triggers phosphorylation cascades through several pathways

Proteins were transferred to PVDF membranes (Thermofisher) using a Power BlotterSemi-dry Transfer System (Thermofisher) and incubated overnight with IFN- (E3V1X) Rabbit mAb (Cell Signaling, #98139S), Galectin-1/LGALS1 (D608T) Rabbit mAb (Cell Signaling, #12936S, Granzyme B (E5V2L) Rabbit mAb (Cell signaling, #44153) and SHMT2 (E7F4Q) Rabbit mAb (Cell signaling, #3344), vinculin Mouse mAb (Sigma-aldrich, #V9131), actin (clone C4) Mouse mAb (MP biomedicals, #691001) antibodies diluted in 5% skimmed milk in Tris-Buffered NaCl Solution with Tween 20 (TBST)

doi: 10.1136/jnnp-2012-304792 166

Reduced glutathione (GSH) reacts with 5,5-Dithiobis (2-nitrobenzoic acid) DTNB in the reaction mix to generate TNB which has an absorbance maximum at 405 nm