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. 2013 Feb 14;8(2):e56891. doi: 10.1371/journal.pone.0056891

Figure 4. ERK1/2-selective inhibitor, U0126, overrides IL-1β- and C/EBPβ-mediated increases in TIMP-1 promoter activity.

Figure 4

Astrocytes were cultured in 48-well plates for 24 h and then cotransfected with TIMP-1 promoter-driven luciferase plasmids (B, −4200/+96 or C −1718/+96) and internal control pGL3-RL. (A) The −4200/+96 promoter harbors an IL-1β responsive element, while the −1718/+96 is the basal TIMP-1 promoter. Both promoters possess a predicted CCAAT-binding site prior to the start site [44]. Cells were allowed to recover for 24 h and then treated with MAPK-selective inhibitors (SB203580, 20 µM; and U0126, 20 µM) for 1 h prior to activation with IL-1β (20 ng/ml) for 24 h. Total cell lysates were collected 24 h post-inhibitor treatment and assayed for luciferase expression. Data represent mean values ± standard error of the mean in at least three independent experiments in at least two independent donors (*p<0.05, **p<0.01, ***p<0.001; significance indicates versus untreated unless indicated by bar).