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. 2016 Jan 28;6:310. doi: 10.3389/fphar.2015.00310

FIGURE 2.

FIGURE 2

Reactive oxygen species (ROS) signal is involved in TNF-α-induced VCAM-1 expression in HCFs. (A) HCFs were pretreated with edaravone (0.1, 1, or 10 μM) for 1 h, and then incubated with TNF-α for 16 h. The protein levels of VCAM-1 were determined by Western blot. (B) HCFs were pretreated with edaravone (10 μM) for 1 h, and then incubated with TNF-α for 4 h. The mRNA expression (white bar) and promoter activity (gray bar) of VCAM-1 were determined by RT/real-time PCR or promoter report assay, respectively. (C) HCFs were incubated with the DCF-DA (5 μM) for 45 min, followed by pretreatment with (or without) edaravone (10 μM) and stimulation with TNF-α for the indicated time intervals or 60 min in the presence of edaravone. The fluorescence intensity of cells was determined. Data are expressed as mean ± SEM of three independent experiments (n = 3). ∗P < 0.05; #P < 0.01, as compared with the cells exposed to TNF-α alone for 60 min.