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. 2015 Jun 8;5:11149. doi: 10.1038/srep11149

Figure 4. Effects of 4VP on cyclins and signaling kinases expressions and the activation of NFκB transcription factor in endothelial cells.

Figure 4

Western blot analyses of effect of 4VP on (A) cyclin B1, cyclin D1, p21 and ERK as well as (B) PI3K, AKT and P38 MAPK expressions. Cells were treated with 4VP (20 or 40 μg/mL) for 24 or 48 hours. Immunoblotting was performed 3–4 times using independently prepared cell lysates and the representative blots were shown. The histograms showed the quantified results of protein levels, which were adjusted with corresponding β-actin protein levels and expressed as fold of control (mean fold of control + SD from 3–4 independent experiments). Differences between the treated and vehicle-treated control groups were determined by one-way ANOVA with Tukey’s post-hoc test. *p < 0.05, **p < 0.01, ***p < 0.005 as compared among groups. (C) Western blot analyses of NFκB expressions in whole cell extract of TNF-α-activated HUVEC and HMEC-1 with or without 4VP. (D) Cells were treated with 4VP (20 or 40 μg/mL) for 6 hours and activated with TNF-α (15 ng/mL). The nuclear extracts were assayed for NFκB activation by NFκB p65 Transcription Factor Assay. Data are representative of two independent experiments.