In this study, we used the Ub fusion model to determine whether wild type mature Smac maintains a proapoptotic function after prolonged cytosolic localization, and evaluate the requirement of the IBM to induce apoptosis [37], [38], [39].
We have identified an interesting OXPHOS-dependent apoptotic mechanism via mtDNA upregulation, and therefore, have evaluated the requirement of this pathway for the efficacy of a wide array of anticancer agents.
However, these studies only evaluated the requirement of single genes for resistance to DNA-damaging agents, whereas the effects on genetic networks were only studied in a small directed screen (St Onge et al. 2007).
To evaluate the requirement of phagocytosis and cathepsin B release in oA β-induced IL-1 β secretion, phagocytosis and cathepsin B were pharmacologically inhibited with cytochalasin D and cathepsin B inhibitor, respectively.
The majority of these factors had been identified in a prior screen that evaluated the requirement of chromatin regulators in MLL-AF9/NrasG12D acute myeloid leukemia (AML) (Zuber et al., 2011b), however TRIM33 was identified here as a unique requirement in B-ALL.
Analysis of gene expression in the presence of GR and ER inhibitors indicates that these receptors are required for the antagonistic regulation of CA12, LEFTY1, and GILZ by Dex and Gen. To evaluate the requirement of GR and ERα more specifically, cells were transfected with siRNA against GR and ERα prior to agonist treatment.
Our understanding of these mechanisms is built predominantly on cell-culture studies of various cell types, although, additionally, animal models have been used to evaluate the requirement of specific proteins in IL-1β-mediated inflammation (Horai et al., 2000; Kuida et al., 1995).
