Fig. 6
IFN-α induces cell type-specific phosphorylation of JAK/STAT pathway components in treated microglia and astrocytes. A IFN-α-associated JAK/STAT signaling pathway depicting phosphosites and their log2 fold change at 5, 15 and 30 min compared with 0 min. Phosphosites were included if they were significantly regulated at one timepoint. Grey boxes: did not reach significance. B Immunoblot of whole protein lysates from IFN-α-treated microglia and astrocytes (n = 3 per cell type per timepoint). Sample “L” was a cross-membrane loading control of a pooled protein from all 30 min IFN-α-treated microglia and astrocyte samples. C Densitometric quantifications of immunoblots. Mean ± SEM are shown. * P < 0.05, ** P < 0.01, *** P < 0.001 and **** P < 0.0001 compared to the respective 0 min of the cell type or between indicated samples as determined by two-way ANOVA with Tukey’s post-test. D Similar trends in fold change between phosphoproteomics and immunoblots. Fold changes calculated from immunoblots are based on the mean intensities at 0 min of treatment and significance determined by two-way ANOVA with Tukey’s post-test. Significance set at |z-score|≥ 1 for phosphoproteomics and P < 0.05 for immunoblots. Dashed horizontal line indicates log2 of a 1.5-fold change. n.d.: not detected