Figure 5.

Overexpression of ATP13A4 WT induces JNK phosphorylation in MCF10A cells. MCF7, non-transduced (NTS) MCF10A cells and MCF10A cells overexpressing ATP13A4 WT (WT OE) or a catalytically dead mutant (D486N OE) were treated for indicated times with 100 µM spermidine (A,B) or 100 µM spermine (C,D). Expression levels of phospho-JNK were determined by Western blot analysis. (A,C) depict representative blots of at least three independent experiments. Of note, the blots are overexposed to show the presence of phospho-JNK bands in MCF7 cells. (B,D) show the bar graphs of the quantification of phospho-JNK levels. Data were normalized to MCF10A WT OE, an untreated control, and are presented as the mean ± s.e.m. Statistical significance was determined by two-way ANOVA with Dunnett’s multiple comparisons test, comparing the different time points to untreated controls for each cell line. p values are depicted in the graphs; non-significant values are not indicated.