Figure 5.
MondoA and MYC gene expression inversely correlates, MondoA enhances pharmacological MYC inhibition, and loss of MondoA leads to significant redistribution of MYC toward DNA-binding sites shared with MondoA. (A) MYC (202431_s_at) expression in different subtypes of B-ALL defined according to chromosomal aberration, normal B cells, and whole blood. (B) MondoA (MLXIP, 202519_at) expression in different subtypes of B-ALL defined according to chromosomal aberration. All data sets were normalized simultaneously using RMA and custom microarray (v15 ENTREZG) CDF files. (C) Dot plot indicates significant inverse correlation between MYC and MondoA (MLXIP) gene expression. RNA-Seq data from TARGET Pan-Cancer (Xena). (D) WB showing MondoA, poly (ADP-ribose) polymerase (PARP), cleaved-PARP (cl-PARP), and MYC under pharmacological inhibition of MYC by JQ1 (400 nM, 16 hours), GAPDH as loading control. (E) Line graph showing relative cell viability after 48 hours of JQ1 treatment at various concentrations. The y-axis represents the percentage of viable cells (Ctrl, MKO, and MOE cells) related to total number of cells as measured by  Celigo S Imaging Cytometer (Nexcelom Bioscience). Live cells were measured by costaining cells with Hoechst 33342 to identify nuclei and fluorescently labeled CalceinAM (C3099; Thermo Fisher Scientific) to identify live cells. (F) Overlap of MondoA-bound genes with genes that are differentially expressed in MKO (log FC, ≥0.4 and ≤ −0.4). The diagram created with online tool Venny 2.1. Detailed ChIP-Seq data in supplemental Table 1. MondoA binding peaks and joint expression pathway analyses (G) Overlap of MondoA-bound genes with genes that are MYC target genes. (H) Average coverage profile of MYC-binding sites on DNA not overlapping (left) and overlapping with MondoA (right) binding sites. The drop in binding affinity in Myc-MKO compared with Myc-MondoA wildtype (MWT) is noticeably larger in MYC-binding sites, which are distant from MondoA-binding sites. (I) Box plot shows statistically significant increase in log2 Fc of Myc-MKO over Myc-MWT for Myc-binding sites overlapping or in close vicinity to MondoA-binding sites. (J) ChIP-Seq peak coverage was visualized with Integrated Genome Browser (IGB). Gene description is given in “Results.” cALL, common ALL. **P < .01; ***P < .001.

MondoA and MYC gene expression inversely correlates, MondoA enhances pharmacological MYC inhibition, and loss of MondoA leads to significant redistribution of MYC toward DNA-binding sites shared with MondoA. (A) MYC (202431_s_at) expression in different subtypes of B-ALL defined according to chromosomal aberration, normal B cells, and whole blood. (B) MondoA (MLXIP, 202519_at) expression in different subtypes of B-ALL defined according to chromosomal aberration. All data sets were normalized simultaneously using RMA and custom microarray (v15 ENTREZG) CDF files. (C) Dot plot indicates significant inverse correlation between MYC and MondoA (MLXIP) gene expression. RNA-Seq data from TARGET Pan-Cancer (Xena). (D) WB showing MondoA, poly (ADP-ribose) polymerase (PARP), cleaved-PARP (cl-PARP), and MYC under pharmacological inhibition of MYC by JQ1 (400 nM, 16 hours), GAPDH as loading control. (E) Line graph showing relative cell viability after 48 hours of JQ1 treatment at various concentrations. The y-axis represents the percentage of viable cells (Ctrl, MKO, and MOE cells) related to total number of cells as measured by  Celigo S Imaging Cytometer (Nexcelom Bioscience). Live cells were measured by costaining cells with Hoechst 33342 to identify nuclei and fluorescently labeled CalceinAM (C3099; Thermo Fisher Scientific) to identify live cells. (F) Overlap of MondoA-bound genes with genes that are differentially expressed in MKO (log FC, ≥0.4 and ≤ −0.4). The diagram created with online tool Venny 2.1. Detailed ChIP-Seq data in supplemental Table 1. MondoA binding peaks and joint expression pathway analyses (G) Overlap of MondoA-bound genes with genes that are MYC target genes. (H) Average coverage profile of MYC-binding sites on DNA not overlapping (left) and overlapping with MondoA (right) binding sites. The drop in binding affinity in Myc-MKO compared with Myc-MondoA wildtype (MWT) is noticeably larger in MYC-binding sites, which are distant from MondoA-binding sites. (I) Box plot shows statistically significant increase in log2 Fc of Myc-MKO over Myc-MWT for Myc-binding sites overlapping or in close vicinity to MondoA-binding sites. (J) ChIP-Seq peak coverage was visualized with Integrated Genome Browser (IGB). Gene description is given in “Results.” cALL, common ALL. **P < .01; ***P < .001.

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