Evaluation of IPA-predicted regulatory pathways and changes in protein production. Purified monocytes were stimulated in vitro for 5 days. (A) Fold increase in IL-6, IL-10, PGE2, and IL-12p70 in culture supernatants on day 5 (mean ± standard deviation [SD] of 7-8 independently studied donors). For unstimulated cultures, concentrations were: 45 pg/mL of PGE2 and 50 pg/mL of IL-12p70. Because IL-6 and IL-10 could not be detected in unstimulated cultures, the fold increase was calculated based on the sensitivity of the enzyme-linked immunosorbent assay, 10 pg/mL for IL-6 and 30 pg/mL for IL-10. (B-C) Effect of blocking NF-κB (2.5 µM of celastrol), Akt (5 µM of perifosine), p38 MAPK (10 µM of SB203580), ERK (1 µM of PD98059), JNK (10 µM of SP600125), PTGS2 (10 µM of celecoxib), or PTGS1 (50 nM of FR122047) on the generation of macrophages by including inhibitors during culture (mean ± SD of 4-6 independently studied donors per group). *P < .05, **P < .01, ***P < .001 vs unstimulated/control cultures. n.s., not significant.