Journal: Cell

Article Title: CBFβ-SMMHC inhibition triggers apoptosis by disrupting MYC chromatin dynamics in acute myeloid leukemia

doi: 10.1016/j.cell.2018.05.048

Figure Lengend Snippet: (A) Analysis of ATAC-seq and K3K27ac and RUNX1 ChIP-seq profiles in a 2 Mb genomic region downstream of MYC. The three enhancer regions (ME1, ME2 and BDME) are depicted in green. (B-D) ChIP-qPCR analysis for RUNX1 in DMSO or AI-10–49 treated cells (B), in DMSO or AI-10–49 treated human primary CD34+ inv(16) AML cells (C), and for p300 in DMSO or AI-10–49 treated ME1 (D). (E) 5C interaction matrices for the MYC locus (−1Mb to +3Mb) in DMSO (left panel) and AI-10–49 (middle panel) treated ME-1 cells. The right panel shows the log2(AI-10–49/DMSO) ratio of the interaction matrices (blue color scheme: higher interaction frequencies in DMSO treated cells; orange color scheme: higher interaction frequencies in AI-10–49 treated cells). Arrows indicate TAD boundaries, arrowhead points to an example of a CTCF-CTCF looping interaction. Significance was calculated using unpaired t-test, *P < 0.05, or **P < 0.005. See also Figures S5 and S6.

Article Snippet: Primary Hematopoietic Cell Cultures Human cord blood samples were collected from the UMASS Memorial Medical Center, and CD34 + cells were isolated using the CD34 Microbead kit (Miltenyi Biotec).

Techniques: ChIP-sequencing, ChIP-qPCR

Journal: Cell

Article Title: CBFβ-SMMHC inhibition triggers apoptosis by disrupting MYC chromatin dynamics in acute myeloid leukemia

doi: 10.1016/j.cell.2018.05.048

Figure Lengend Snippet: (A) Heat map representation of differentially expressed genes in RNA-seq analysis between DMSO and AI-10–49 treated ME-1 cells. A total of 591 genes (in red) are up- and 696 genes (in blue) down-regulated in AI-10–49 treated cells (>2fold change, FDR<0.01). (B) Gene set enrichment analysis showing biological processes and signaling pathways correlated with AI-10–49-treatment. (FDR) false discovery rate; (NES) normalized enrichment score. (C) MYC transcript levels in ME-1 cells treated with DMSO (D) or AI-10–49 (49), estimated by RT-PCR. (D) MYC, ERK and LMO1 nascent transcript levels in treated ME-1 cells, estimated by transient transcriptome qRT-PCR. (E) MYC protein levels in treated ME-1 cells, by immunoblotting. (F, G) MYC transcriptional levels in mouse lineage negative wild type bone marrow (mu wt Lin- BM) and lineage negative CBFβ+/MYH11 leukemic cells (F), and human CD34+ cord blood (HPCs) and human primary CD34+ inv(16) AML cells (G); each symbol represents the average of a triplicate experiment, and the average value of three samples is shown in red. Experiments in A, C and D were performed in triplicate treatments. Error bars represent the SD. Significance was calculated using unpaired t-test (C, D, F and G), *P < 0.05, or **P < 0.005. See also Figure S1.

Article Snippet: Primary Hematopoietic Cell Cultures Human cord blood samples were collected from the UMASS Memorial Medical Center, and CD34 + cells were isolated using the CD34 Microbead kit (Miltenyi Biotec).

Techniques: RNA Sequencing, Protein-Protein interactions, Reverse Transcription Polymerase Chain Reaction, Quantitative RT-PCR, Western Blot

Journal: Cell

Article Title: CBFβ-SMMHC inhibition triggers apoptosis by disrupting MYC chromatin dynamics in acute myeloid leukemia

doi: 10.1016/j.cell.2018.05.048

Figure Lengend Snippet: KEY RESOURCES TABLE

Article Snippet: Primary Hematopoietic Cell Cultures Human cord blood samples were collected from the UMASS Memorial Medical Center, and CD34 + cells were isolated using the CD34 Microbead kit (Miltenyi Biotec).

Techniques: Recombinant, SYBR Green Assay, Extraction, Proliferation Assay, Sample Prep, shRNA, Plasmid Preparation, Cloning, Software, Red Blood Cell Lysis, Transfection, Protease Inhibitor, Gel Extraction