Fig. 7.

Abolishment of the effect of IFNβ on NCC migration by inhibition of the JAK-STAT pathway. a, b NCC were pre-treated for 0.5 h with two different JAK inhibitors (ruxolitinib and tofacitinib) at the indicated concentrations. Then, the cells were further treated for 1 h with the inhibitors in cell culture medium supplemented with 500 pM IFNβ. Finally, cells were harvested and the amount of phosphorylated STAT1 (p-STAT1) was measured by western blot analysis (representative blots are shown). c NCC were exposed to IFNβ for 48 h at the indicated concentrations, either with or without ruxolitinib (Rux, 10 µM). Then, cells were harvested, and total RNA was extracted and retro-transcribed. Effects on selected mRNAs were evaluated by qPCR. Expression levels were normalized against the housekeeping gene, GAPDH and are expressed relative to control levels (untreated cells). The mRNA expression in the presence of Rux is shown in red. Note that the red symbols are often overlapping, due to complete inhibition down to control levels. d, e Band intensities were quantified for p-STAT1 (normalized to GAPDH). In a parallel set of experiments, migration (after 48 h) was evaluated in the presence of IFNβ (500 pM) plus ruxolitinib (left) or tofacitinib (right). Data are from three independent experiments. Error bars indicate standard deviations (SD). Statistics was performed by ANOVA, followed by Dunnet’s post-hoc test (*p ≤ 0.05, **p ≤ 0.01, ***p ≤ 0.001). (Color figure online)