<i>Twist2</i> does not impair the expression of clock genes.

<p>(A) Expression of <i>Twist2</i> mRNA in NIH-3T3 cells after a 4 hr treatment with TNF, IL-1β and TGFβ2. The concentrations of the cytokines used are given in the legend to <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0137229#pone.0137229.g001" target="_blank">Fig 1</a>. Data of RT-qPCR assays show the mean +/- SEM of three independent experiments. (B) Western blot showing upregulation of TWIST2 in cell lysates of NIH-3T3 cells treated with TNF for 4 hr. Nuclear matrix protein p84 was used as loading control. (C) Transfection of NIH-3T3 cells with pCMV6::<i>Twist2</i> show no significant effect on expression of <i>Dbp</i>, <i>Per1</i>, <i>Per2</i> and <i>Per3</i> compared to the empty vector control. (D) Data show TWIST2 expression in untreated (empty vector control) and TNF treated NIH-3T3 cells and in cells transfected with pCMV6::<i>Twist2</i>. Data of RT-qPCR assays show the mean +/- SEM (error bars) of triplicates from three independent experiments. Significance was calculated with two-way ANOVA and Bonferroni post-hoc test; *** p<0.001. Western blots show one representative experiment of three.