TBHQ (tert-Butylhydroquinone) is an antioxidant that induces an antioxidant response through the redox-sensitive transcription factor Nrf2.

Jurkat T cells were cultured in 96-well plates (1×10^5 cells/well) and treated as described, and cell supernatants were harvested 24h after activation with anti-CD3/anti-CD28. IL-2 was quantified from cell supernatants by the use of a commercially available kit per the manufacturer's protocol. Relative light intensity was quantified at 450nm by Bio-Tek μQuant microplate reader. The IL-2 concentrations of each sample were then calculated using a line ar standard curve on Microsoft Excel [2].

TBHQ powder was directly mixed with the animal food powder at a ratio of 1% (w/w) as described previously. The efficacy of this protocol for TBHQ treatment in vivo has also been validated by other groups. TBHQ treatment was continued for 4 weeks after the TAC surgery [3].