Fig. 1.

IFNa effects and mRNA regulation in 32D-BCR-ABL- and 32D-JAK2V617F-positive cells. 32D-JAK2V617F-positive cells showed concentration-dependent decreased viability after 72 h of IFNa treatment and activation of interferon target gene expression compared to BCR-ABL cells. a 32D-EV (black), 32D-BCR-ABL (blue), and 32D-JAK2V617F (red) were treated with IFNa (0–10⁴ U/ml) for 72 h, and the viability was measured by MTT. 32D-EV cells were grown in the presence of 5 ng/ml IL-3. b Gene set enrichment analysis of ISG and IL2/STAT5 genes. c Microarray results were validated by RT-qPCR. mRNA expression of EV-positive (green), BCR-ABL-positive (blue), or JAK2V617F-positive (red) cells was analyzed in murine 32D cells, which were WEHI starved for 24 h, or in human cell lines harboring the same genetic alteration. Oncogene-transduced cells were treated with 1 μM TKI (BCR-ABL + imatinib; JAK2V617F + ruxolitinib) for 18 h prior to analysis. The expression level was calculated as a percentage of Gapdh. Depicted are the mean values ± SD. *p < 0.05, **p < 0.01, ***p < 0.001. d Mononucleated cells were isolated from peripheral blood samples of healthy volunteers (green), BCR-ABL-positive (blue) CML patients, and JAK2V617F positive (red) PV patients, and the mRNA expression was analyzed as a percentage of GAPDH and analyzed with one-way ANOVA and Dunn’s multiple comparison test. STAT1 and IRF1: controls: n = 16, CML: n = 13, PV: n = 8; STAT2: controls: n = 5, CML: n = 8, PV: n = 6. IRF9: controls: n = 7, CML: n = 9, PV: n = 6. e Gene expression microarray analysis of CD34-positive cells isolated from BCR-ABL-negative MPN or BCR-ABL-positive CML patients. Fold change of gene expression is shown, depicting downregulation of the analyzed gene in blue and upregulation in red