Interleukin-4 in the Generation of the AERD Phenotype: Implications for Molecular Mechanisms Driving Therapeutic Benefit of Aspirin Desensitization
EMSA for STAT6. (a) EMSAs were performed using 32P-labeled oligomers comprising the STAT6 site within the CysLT1R promoter. Nuclear extracts were purified from THP-1 mononuclear cell lines in the resting state, IL-4 stimulated (10 ng/mL), and IL-4 stimulated in the additional presence of aspirin (10 mM). STAT6 binding was evaluated by performing EMSAs in the presence of 100–300-fold molar excess unlabeled STAT6 consensus sequence (comprising the ε heavy chain promoter) or a mutated STAT6 consensus sequence. EMSAs were also performed using STAT6, phosphoSTAT6, and, as a control, STAT4 antibodies. (b) Relevance to normal tissue was evaluated using nuclear extracts prepared as above, derived from enriched peripheral blood-derived mononuclear phagocytes .
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