Figure 6.
Transcriptome analysis of M/E p53WT AML treated with CX-5461 by RNA sequencing revealed induction of myeloid maturation. (A) Schematic diagram of the experimental design (n = 3 mice per group). M/E p53WT AML were treated with a single dose of CX-5461 (40 mg/kg). (B) Scatter-plots for p53WT M/E illustrating logFC over CPM at 10 hours, and (C) GeneGo pathway analysis. (D) M/E p53WT or p53null AML were engrafted into recipient C57Bl/6 mice and treated with CX-5461 for 48 hours. Differential cell classification was performed on May-Grünwald Giemsa-stained cytospins prepared from GFP+-sorted BM (****P < .0001, ***P = .0008, **P = .0066; n.s. P = .0036; n = 5). Graphs represent mean ± SEM. Insert is a representative image from 1 mouse per group. Scale bar represents 10 μm. (E) Expression of Mac-1, as determined by flow cytometry, in the BM-derived tumor cells (M/E p53WT and p53null, MFI, ****P < .0001; n = 5). (F) Representative flow cytometry dot plots from 1 mouse per group and quantitation of c-Kit expression in GFP+ tumor cells from the BM of M/E p53WT leukemic mice (****P < .0001; n = 5). (G) Mac-1 cell surface expression in KG-1, THP-1, ML-2, and MOLM-13 cells in response to CX-5461 or CX-5447 treatment after 24 hours (n = 3). Graph represents mean ± SEM. Unpaired 2-tailed Student t test (A,D) was used. CPM, counts per million; CRTH2, chemoattractant receptor-homologous molecule expressed on T helper type 2 cells; FACS, fluorescence-activated cell sorting; FDR, false discovery rate; G-CSF, granulocyte colony-stimulating factor; logFC, log fold-change; MFI, mean fluorescence intensity; n.s., not significant; PTAFR, platelet-activating factor receptor; RNA-seq, RNA sequencing; Th2, T helper 2.