Research Paper Volume 13, Issue 8 pp 11969—11987

Figure 7. CCL2 induced FoxO1 activation in a p38 MAPK-dependent manner. (A and B) THP-1 was incubated with rCCL2 (50 ng/mL) for the indicated time intervals or pretreated with 10 μM SB203580, 10 μM SP600125, or 50 nM SCH772984 for 24 h, and then incubated with rCCL2 (50 ng/mL) for 120 min. The ChIP assay was applied to examine the enrichment of FoxO1 onto the COX-2 promoter. (C and D) THP-1 cells were transfected with FoxO1 expression vectors (50 ng), and pGL3-COX-2 (COX-2 promoter-driven luciferase) construct (200 ng) plus pRL-TK Renilla (10 ng). After 36 h, the cells were treated with or without AS1842856 (1 μM) or plus 10 μM SB203580, 10 μM SP600125, or 50 nM SCH772984 for a further 24 h. (E) THP-1 cells were co-transfected with a wild-type COX-2 promoter reporter gene (Luc-COX-2-WT) or FoxO1 binding site-mutated COX-2 promoter reporter gene (Luc-COX-2-mut) with FoxO1. The COX-2 promoter activity was determined. Luciferase activity was normalized with Renilla luciferase values. Data are expressed as fold change relative to the level of control. Data are presented as mean ± standard error. ^*, P < 0.05.