Macrophage activation in response to A2t involves both MyD88 and TRIF. (A) WT or MyD88^−/− 7-day-old murine BMDM were treated with 5nM A2t for 24 hours, and levels of the indicated cytokines (in pg/mL) were measured in the supernatants by luminex. Macrophages were pooled from 4 males per genotype in each of 2 separate experiments run in duplicate. Results expressed as the mean ± SEM. (B) Nuclear extracts of 7-day-old human MDM treated with either LPS (2 ng/mL) or A2t (5nM) were tested for p65 or IRF3 activity by binding of plated DNA oligos bearing the specific NF-κB or IRF3 consensus site, followed by colorimetric detection of p65 or IRF3 by horseradish peroxidase–linked antibody.