Phosphorylated AMPK Quantification
SECONDARY
Banked PBMC samples were stained with a fixable viability dye (Zombie NIR, Invitrogen) and a cocktail of surface markers to identify T-cell populations. After surface stain, cells were fixed and permeabilized using the Foxp3 Transcription Factor Staining Buffer Set (eBioscience). Cells were then stained intracellularly using a polyclonal antibody against phosphorylated AMPK (p-AMPK) alpha-1/2 (Thr172, Thr183; Thermo Fisher Scientific) followed by an Alexa488-conjugated secondary antibody. Stained cells were acquired on a 5-laser Cytek Aurora and analyzed using FlowJo software (FlowJo, LLC).The data generated in this study are available upon request from the corresponding author.