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: To analyze the cytotoxicity in the different fractions of the bulk treated AML cells, treated MNCs were blocked with human FcR blocking reagent (Miltenyi Biotec) for 5 min and stained with the following antibodies: CD45-APC-Cy7, TMRE, CD14-PerCP, CD34-PE-Cy7 (or CD117-PE-Cy7 for CD34 − samples), and CD11b-APC for 20 min at 4°C.

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

Journal: bioRxiv

Article Title: Enhanced FLI1 accessibility mediates STAG2-mutant leukemogenesis

doi: 10.1101/2025.04.01.646632

Figure Lengend Snippet: A ) Schematic of experimental set up. B ) The bone marrow counts of Ubc -CreER T2 positive WT (control), Stag2 Δ (single mutant), Npm1 c/+ (single mutant) and Stag2 Δ Npm1 c/+ (double mutant) mice at 4 weeks post mutation activation. Data presented as mean+s.e.m, *p <0.05, One-way ANOVA with Tukey’s multiple comparison. C ) The percentage of LSK (Lineage-Sca1+cKit+) cells in the bone marrow. Data presented as mean+s.e.m, *p <0.05, **p <0.01, ***p <0.001, One-way ANOVA with Tukey’s multiple comparison. D ) The percentage of long-term hematopoietic stem cells (HSCLT, LSKFlk2-Cd150+Cd48-), short-term hematopoietic stem cells (HSCST, LSKFlk2-Cd150-Cd48-), multipotent progenitor 2 (MPP2, LSKFlk2-Cd150+Cd48+), multipotent progenitor 3 (MPP3, LSKFlk2-Cd150-Cd48+), and multipotent progenitor 4 (MPP4, LSKFlk2+Cd150-) cells under the LSK population. ***p <0.001, ****p < 0.0001, Two-way ANOVA with Tukey’s multiple comparison. E ) The percentage of granulocytic-macrophage progenitor (GMP, Lineage-cKit+Sca1-Cd34+FcyR+). Data presented as mean + s.e.m. F-H ) The percentage of mature myeloid (Gr1+Mac1+, F), B lymphoid (B220+Cd19+, G) and immature erythroid (Ter119+Cd71+, H) population in the bone marrow. Data presented as mean + s.e.m, ***p < 0.001, One-way ANOVA with Tukey’s multiple comparison. I ) Schematic of LSK transplantation experiment. (n=5-8 biological replicates with n= 1-2 recipients). J ) The chimerism of transplant recipients of LSK cells in the peripheral blood over 16 weeks post transplantation. Data presented as mean+s.e.m, *p <0.05, **p <0.01, Mixed model ANOVA with multiple comparison. K ) The percentage of myeloid (Gr1+Mac1+) population in the donor fraction (Cd45.2+) of the recipients. Bar represents mean+s.e.m, *p <0.05, **p <0.01, Mixed model ANOVA with multiple comparison. L ) The percentage of donor chimerism (Cd45.2%) in the LSK fraction and the peripheral blood (PB). Bar represents mean+s.e.m, *p <0.05, **p <0.01, One-way ANOVA with Tukey’s multiple comparison.

Article Snippet: Blast populations were identified by CD45 and side scatter characteristics and the following antibodies were used to characterize blasts (all obtained from Becton Dickinson): CD45-APC/Cy7, CD34-APC, CD13-PE/Cy7, HLADR-PerCP/Cy5.5, CD33-PE, and CD65-FITC.

Techniques: Control, Mutagenesis, Activation Assay, Comparison, Transplantation Assay