2.5. Transfection Experiments

To evaluate the effect of PML/RARa on NRF2 half life, Hek293T cells were transfected with pSG5-PML/RARa or pSG5 constructs using jetPRIME^®transfection reagent (Polyplus-transfection^®, Strasbourg, France). The cells were treated with ZnSO₄ 100 µM for two hours to induce NRF2 expression and then the stops were initiated over five hours.

HL-60 cells were transfected with pSG5-PML/RARa or pSG5 constructs using Amaxa Cell Line Nucleofector Kit C (Lonza, Cologne GmbH, Germany). After 24 h the cells were treated with ZnSO4 100 µM for two hours to induce NRF2 expression and then with cycloheximide (CHX) (Sigma-Aldrich, St. Louis, MO, USA), 0.2 nM for another four hours.