Journal: Cell Reports Medicine

Article Title: ΔNp73 isoform defines a TP53 -mutant-like poor-risk subgroup of acute myeloid leukemia

doi: 10.1016/j.xcrm.2025.102540

Figure Lengend Snippet: ΔNp73 overexpression is associated with downregulation of the TP53 signaling pathway in TP53 wt AMLs (A) Western blot analysis for ΔNp73 and total TP73 in total cell extracts from MOLM13 cells transduced with lentivirus containing the EV (pMEG) or the ΔNp73α or ΔNp73β cDNA. (B) Volcano plot displaying the differentially expressed genes in MOLM13 cells with ΔNp73-OE versus EV control ( n = 2). (C) Expression of CD14 and CD117 in MOLM13 EV (pMEG) and ΔNp73α-OE cells ( n = 3). (D) GSEA analysis using the fold change values from the analysis depicted in (A). False discovery rate (FDR)-q values are indicated. (E) ChIP-seq data on MOLM13 cells used in (A) using antibodies against TP53 or GFP (for the GFP-ΔNp73 fusion), and TAp73. Heatmaps with signals ± 5 kb from the transcription start site (TSS) are shown. (F) Representative screenshots of TP53, TAp73, and ΔNp73 antibody binding at four TP53 target loci. (G) Venn diagram depicting overlapping peaks detected for the TP53 ChIP-seq in MOLM13 EV control cells and the GFP-ΔNp73 in MOLM13-ΔNp73 OE cells. Lower: GO analysis for the overlapping peaks (51 targets). (H and I) Cumulative cell count of MOLM13 ( TP53 wt, H) and TF1 ( TP53 mut, I) cells transduced with ΔNp73α, ΔNp73β, and EV control, cultured for 9 days ( n = 4). (J) Western blot analysis for TP53 and total TP73 in total cell extracts from MOLM13 cells transduced with EV (pMEG) or the shRNA targeting the TP53 gene (shTP53). Cumulative cell count of MOLM13 TP53 KD cells (sh TP53 ) transduced with ΔNp73α, ΔNp73β, and EV control, cultured for 9 days, is shown in the right ( n = 4). Data are reported as mean ± SEM for (H) and (I). The p values and cell lines are indicated in the graphs; ∗ p < 0.05; ∗∗ p < 0.01; ∗∗∗ p < 0.001; ANOVA and Bonferroni post-test.

Article Snippet: CD117 MicroBeads Kit, Human , Miltenyi Biotec , 130-091-332.

Techniques: Over Expression, Western Blot, Transduction, Control, Expressing, ChIP-sequencing, Binding Assay, Cell Characterization, Cell Culture, shRNA

Journal: Cell Reports Medicine

Article Title: ΔNp73 isoform defines a TP53 -mutant-like poor-risk subgroup of acute myeloid leukemia

doi: 10.1016/j.xcrm.2025.102540

Figure Lengend Snippet: Pharmacological and genetic inhibition of CEBPA synergizes with VEN-induced apoptosis in TP53mut/mut-like AMLs (A) MOLM13 (VEN-sensitive) and KG1 (VEN-resistant) cells were treated for 72 h with increasing concentrations of VEN and GFC. Synergy was determined by Bliss coefficient (ZIP score >10 indicates synergism). (B and C) Drug-induced apoptosis (B) and viable cell counts (C) in MOLM13 shCEBPA/shScr cells treated with VEN alone or in combination with GFC (concentrations indicated in the plots, 72 h) detected by flow cytometry ( n = 4). (D and E) Apoptosis was detected by flow cytometry in gated human CD45 dim CD34 + (or CD117 + cells for CD34 − AMLs) of ex vivo -treated AML samples categorized as TP53 mut-like ( n = 8) (D) and TP53 mut ( n = 9) (E) in a co-culture system using an FITC-annexin V/DAPI staining method. Cells were treated with vehicle, VEN (100 and 500 nM), and VEN+Aza (VEN 100 nM + 5′ Aza 1.5 μM), in the presence or absence of GFC (30 μM) for 72 h. Bar graphs represent the mean ± SEM of all the independent patients screened; each point represents a patient. (F and G) Mitochondrial membrane potential (F) (measured by TMRE staining, n = 18) and total cytoplasmatic ROS levels (G) (measured using the CellROX Red probe, via flow cytometry, n = 6) for the data included in (D) and (E). TP53 mut AMLs are depicted in red, and TP53 mut-like AMLs are depicted in black. APR-246, eprenetapopt. Data are reported as mean ± SEM for (B)–(G). The p values and cell types are indicated in the graphs; ∗ p < 0.05; ∗∗ p < 0.01; ∗∗∗ p < 0.001; ANOVA and Bonferroni post-test.

Article Snippet: CD117 MicroBeads Kit, Human , Miltenyi Biotec , 130-091-332.

Techniques: Inhibition, Flow Cytometry, Ex Vivo, Co-Culture Assay, Staining, Membrane