Fig. 6: Ruxolitinib treated JAK2-V617 display hyperactivation of STAT5 target genes PIM, ID1 and MPL after ruxolitinib dissociation.

Gene set enrichment analysis (GSEA) identifies Hallmark gene sets enriched in Ruxo-treated compared to DMSO-treated samples. The x-axis represents the -log10(p value), indicating the statistical significance of enrichment. The y-axis shows the names of the Hallmark gene sets. The color of each bar represents the direction of enrichment, with green indicating upregulation and red indicating downregulation of genes within the corresponding set (A). Heatmap depicting the log-CPM expression values (Z-score scaled) of selected genes across DMSO-treated and Ruxo-treated experimental conditions. The x-axis represents the experimental groups replicates, and the y-axis shows the individual genes. P value scale at the left provides statistical significance reference for interpreting the expression values (B). Heatmap depicting the log-CPM expression values (Z-score scaled) of selected genes across DMSO-wash and Ruxo-wash experimental conditions. The x-axis represents the experimental groups replicates, and the y-axis shows the individual genes. P value scale at the left provides statistical significance reference for interpreting the expression values (C). Quantification of representative values of PIM2, ID1, MPL and TGM2 was shown between the DMSO wash and ruxolitinib wash samples (D). RNA-seq data of MPN patients treated with ruxolitinib (n = 36) and non-JAK inhibitor treatment (n = 126) was analyzed for PIM2 and MPL transcripts levels (E). Immunoblot analysis of ruxolitinib persisters developed were subjected to indicated antibodies (F). A representative image of n = 2 two independent experiments is shown. ****p < 0.0001; ***p < 0.001, *p < 0.05 and n.s., not significant, p > 0.05 by Student’s t test.