Fig. 1

EP300-ZNF384 promotes the expression of IL3RA in B-cell acute lymphoblastic leukemia (B-ALL). (A) Volcano plot of comparative analysis between EP300-ZNF384 positive (n = 7) and negative (n = 190) patients in the TARGET-ALL-P2 cohort. Upregulated (n = 346) and downregulated (n = 216) genes are colored by red and green (fold change > 2 and P < 0.01). The labeled texts showed the upregulated genes in the cytokine-cytokine receptor interaction pathway. (B) Bar plot of enriched Kyoto Encyclopedia of Genes and Genomes gene sets using the differentially expressed genes (fold change > 2 and P < 0.01) from the comparison of the EP300-ZNF384 positive and negative patients in the TARGET-ALL-P2 cohort. (C) Representative GSEA plot of EP300-ZNF384-positive patients compared with EP300-ZNF384-negative patients. The normalized enrichment score (NES) and nominal P-values are shown in the graph. (D) Heatmap of 12 upregulated genes in the cytokine-cytokine receptor interaction pathway in EP300-ZNF384-positive patients. IL3 and IL3RA are labeled with blue. (E) The empty vector, EP300-ZNF384, and ZNF384 were overexpressed in NALM-6 cells through lentivirus-mediated gene transfer. Puromycin (2 µg/mL)-selected cells were collected for RT-qPCR. The expression of EP300-ZNF384 together with those of 12 upregulated genes in the cytokine and cytokine receptor interaction pathway were determined relative to ACTB expression. Data represent the mean ± SD. *P < 0.05, **P < 0.01, ***P < 0.001. (F) Immunoblotting analysis of the EP300-ZNF384 fusion and wild-type ZNF384 proteins in the empty vector, EP300-ZNF384, and ZNF384-overexpressing NALM-6 cells. (G) Surface expression of CD123 was evaluated by flow cytometry on NALM-6 cells with the empty vector, EP300-ZNF384 fusion gene, and ZNF384. The proportion of CD123-positive cells and the median fluorescence intensity of CD123 were quantified. *P < 0.05, ***P < 0.001