Pharmacological inhibition of JAK2/STAT signaling reduces AURKA and AURKB expression in SET2 cells. (A) Gene expression heatmap from RNA-seq analysis of SET2 cells upon the treatment with ruxolitinib. Data was obtained from Meyer et al.20 (GEO accession GSE69827). Gene expression was expressed as fold change of the mean of normalized counts of naïve SET2 cells, which was set as 1; genes demonstrating ≥1.5-fold in either direction are included in the heatmap. The mean of fold change obtained from three experimental replicates is indicated. (B) qPCR analysis of AURKA and AURKB mRNA expression in SET2 cells treated with graded concentrations of ruxolitinib (vehicle, 100, 300, or 1000 nM) for 48 hours. Bar graphs represent the mean ± SD of at least four independent experiments. The p values are indicated in the graphs; *p < 0.05, ***p < 0.0001; ANOVA test and Bonferroni post-test. (C) Western blot analysis for p-STAT3Y705, p-STAT5Y694, AURKA, AURKB, and p-histone H3S10, in total cell extracts from SET2 cells treated with graded concentrations of ruxolitinib (vehicle, 100, 300, or 1000 nM) for 48 hours (Right panel) or graded time of exposure (0, 3, 6, 9, 12, 24, and 48 hours) to ruxolitinib at 300 nM (Left panel); membranes were reprobed with the antibody for the detection of the respective total protein or α-tubulin, and developed with the SuperSignal™ West Dura Extended Duration Substrate system using a Gel Doc XR+ imaging system. Cropped blots are shown and full-length blots are included in Supplementary Fig. 7.