promyelocytic leukemia cells  (ATCC)

Journal: Blood Neoplasia

Article Title: Acid ceramidase inhibition enhances BCL-2 targeting in venetoclax-resistant acute myeloid leukemia

doi: 10.1016/j.bneo.2026.100196

Figure Lengend Snippet: Pharmacologic inhibition of AC enhanced venetoclax cytotoxicity in venetoclax-resistant AML cell lines. (A) Correlation analyses comparing venetoclax AUC values and ASAH1 mRNA expression (n = 332) from BeatAML2.0 ( http://www.vizome.org/aml2/ ). Dotted lines represent linear regression with 95% confidence interval. (B) MV4-11 parental and MV4-11 VEN-R cells were subjected to increasing concentrations of venetoclax for 48 hours, and cell viability was assessed by MTS. Dotted lines represent 95% confidence interval. (C) Immunoblotting of MV4-11 parental and MV4-11 VEN-R cells. β-Actin expression served as the loading control for immunoblots, and a representative blot is shown. Protein levels were quantified relative to β-actin and normalized to parental. (D-E) MTS cell viability, flow cytometry annexin V staining, and Bliss synergy scores (SynergyFinder 2.0) were measured following dimethyl sulfoxide (DMSO), SACLAC, venetoclax, or combination treatment at the indicated concentrations in MM-6 (48 hours) and MV4-11 VEN-R cells (72 hours). Bliss synergy scores were calculated for each drug combination tested, and the average Bliss score is shown. Bliss scores >10 were considered synergistic, between −10 and 10 were considered additive, and less than >10 were considered antagonistic. (F) MM-6 and MV4-11 VEN-R cells were treated with DMSO, SACLAC, venetoclax, or the combination at the indicated concentrations for 48 hours or 72 hours, respectively, and annexin V and 7-AAD population percentages were measured by flow cytometry. Significance was assessed by 2-way analysis of variance (ANOVA) with Tukey multiple comparisons test comparing live cells (annexin negative, 7-AAD negative) across treatment groups. Quantitative data are presented as mean ± standard deviation (SD). Results are representative from 2 independent experiments (panels B-C) and the average of 3 independent experiments (panels D-F). Concentrations in brackets are in micromolar. ∗∗∗∗ P < .0001. ABT-199, venetoclax (VEN); AUC, area under the curve; ns, not significant.

Article Snippet: HL-60 and MV4-11 cells were obtained from ATCC (Manassas, VA).

Techniques: Inhibition, Expressing, Western Blot, Control, Flow Cytometry, Staining, Standard Deviation

Journal: Blood Neoplasia

Article Title: Acid ceramidase inhibition enhances BCL-2 targeting in venetoclax-resistant acute myeloid leukemia

doi: 10.1016/j.bneo.2026.100196

Figure Lengend Snippet: S ACLAC and venetoclax induced ceramide accumulation and caspase-mediated cell death characterized by NOXA accumulation. (A) MM-6 and MV4-11 VEN-R cells were treated with DMSO, 15μM SACLAC, 3μM venetoclax, or the combination for 24 hours, and ceramide levels were measured by liquid chromatography-mass spectrometry. Significance was assessed by 2-way ANOVA with Tukey multiple comparisons test. (B-C) Flow cytometry annexin V and 7-AAD profiling and immunoblotting of MM-6 and MV4-11 VEN-R cells treated with DMSO, 15μM SACLAC, 3μM venetoclax, or the combination at the indicated concentrations for 12, 24, 48, and 72 hours. Significance was assessed by 2-way ANOVA with Tukey multiple comparisons test comparing live cells (annexin negative, 7-AAD negative) across treatment groups. β-Actin expression served as the loading control for immunoblots, and a representative blot is shown. Protein levels were quantified relative to β-actin and normalized to the first lane. (D) Flow cytometry annexin V profiling of cells pretreated with DMSO or 50μM Z-VAD-FMK for 2 hours followed by DMSO, 15μM SACLAC, 3μM venetoclax, or the combination treatment for 48 hours. Significance was assessed by unpaired t test with Welch correction. (E) Immunoblotting of MM-6 cells pretreated with DMSO or 50μM Z-VAD-FMK for 2 hours followed by DMSO, 15μM SACLAC, 3μM venetoclax, or the combination treatment for 48 hours. β-Actin expression served as the loading control for immunoblots, and a representative blot is shown. Cleaved PARP and cleaved caspase-3 levels were quantified relative to β-actin and normalized to the first lane. (F-G) Flow cytometry annexin V profiling and immunoblotting of cells transfected with control small interfering (si)RNA or si PMAIP1 for 24 hours followed by DMSO, 15μM SACLAC, 3μM venetoclax, or the combination treatment for 48 hours. Significance was assessed by unpaired t test with Welch correction. β-Actin expression served as the loading control for immunoblots, and a representative blot is shown. Protein levels were quantified relative to β-actin and normalized to the first lane. Quantitative data are presented as mean ± SD. Results are representative from 2 to 3 independent experiments (panels A,C-G), and the average of 3 independent experiments (panel B). Concentrations in brackets are in micromolar. ∗ P < .05; ∗∗ P < .01; ∗∗∗ P < .005; ∗∗∗∗ P < .0001. ns, not significant; SAC, SACLAC; VEN, venetoclax; Z-VAD, Z-VAD-FMK.

Article Snippet: HL-60 and MV4-11 cells were obtained from ATCC (Manassas, VA).

Techniques: Liquid Chromatography, Mass Spectrometry, Flow Cytometry, Western Blot, Expressing, Control, Transfection

Journal: Blood Neoplasia

Article Title: Acid ceramidase inhibition enhances BCL-2 targeting in venetoclax-resistant acute myeloid leukemia

doi: 10.1016/j.bneo.2026.100196

Figure Lengend Snippet: SACLAC and venetoclax upregulated NOXA via a cytotoxic ISR. (A) Volcano plot of significantly enriched (n = 249) or depleted (n = 190) proteins from SACLAC vs DMSO-treated MV4-11 cells. Significance was defined using an FDR (q) <0.1 cutoff. (B) Biological Process (Gene Ontology) analysis of significantly downregulated proteins from panel A. (C) Immunoblotting of cells treated with DMSO, 15μM SACLAC, 3μM venetoclax, or the combination for 12 or 24 hours. β-Actin expression served as the loading control for immunoblots, and a representative blot is shown. Protein levels were quantified relative to total eIF2α (for p-eIF2α [S51]) or β-actin (for ATF4) and normalized to the first lane. (D) Flow cytometry annexin V profiling of cells pretreated with DMSO or 3μM ISRIB for 2 hours followed by DMSO, 15μM SACLAC, 3μM venetoclax, or the combination treatment for 48 hours. Significance was assessed by unpaired t test with Welch correction. (E) Immunoblotting of cells pretreated with DMSO or 3μM ISRIB for 2 hours followed by DMSO, 15μM SACLAC, 3μM venetoclax, or combination treatment for 24 or 48 hours. β-Actin expression served as the loading control for immunoblots, and a representative blot is shown. Protein levels were quantified relative to total eIF2α (for p-eIF2α [S51]) or β-actin (for ATF4 and NOXA) and normalized to the first lane. (F) Immunoblotting of cells pretreated with DMSO, PKR-IN-C16 (PKR inhibitor), AMG PERK 44 (PERK inhibitor), or GCN2-IN-1 (GCN2 inhibitor) as indicated for 2 hours followed by DMSO or 15μM SACLAC treatment for 24 hours. β-Actin expression served as the loading control for immunoblots, and a representative blot is shown. Protein levels were quantified relative to β-actin and normalized to the first lane. (G) Flow cytometry annexin V profiling of cells pretreated with DMSO or 2.5μM to 5μM GCN2iB (GCN2 inhibitor) for 2 hours followed by DMSO, SACLAC (MM-6 = 15μM; MV4-11 VEN-R = 10μM), venetoclax (MM-6 = 3μM; MV4-11 VEN-R = 1μM), or the combination treatment for 24 hours. Significance was assessed by unpaired t test with Welch correction. Quantitative data are presented as mean ± SD. Results are representative from 2 to 3 independent experiments. Concentrations in brackets are in micromolar. ∗ P < .05; ∗∗ P < .01; ∗∗∗ P < .005. FDR, false discovery rate; ns, not significant; SAC, SACLAC; VEN, venetoclax.

Article Snippet: HL-60 and MV4-11 cells were obtained from ATCC (Manassas, VA).

Techniques: Western Blot, Expressing, Control, Flow Cytometry

Journal: Blood Neoplasia

Article Title: Acid ceramidase inhibition enhances BCL-2 targeting in venetoclax-resistant acute myeloid leukemia

doi: 10.1016/j.bneo.2026.100196

Figure Lengend Snippet: Effect of AC and BCL-2 inhibition on mitochondrial function and membrane potential. (A-B) MM-6 and MV4-11 VEN-R cells were pretreated with DMSO, 50μM Z-VAD-FMK, or 3μM ISRIB for 2 hours, then treated with DMSO, 15μM SACLAC, 3μM venetoclax, or the combination for 48 hours. Mitochondrial membrane potential was assessed by flow cytometry using tetramethylrhodamine, methyl ester. Significance was assessed by unpaired t test with Welch correction. (C-D) MM-6 and MV4-11 VEN-R cells were treated with DMSO, SACLAC, LCL-805, venetoclax, SACLAC + venetoclax, or LCL-805 + venetoclax as indicated. Oxygen consumption was measured in digitonin (0.02 mg/mL) permeabilized cells in the absence of substrates (basal), with ATP free energy clamped at −54 kJ/mol (ΔGATP −54 ), or following the addition of complex I substrates (Oct-Carn, Glut), complex II substrates (Succ), the respiratory uncoupler FC, or the complex I inhibitor rotenone. Note different y-axis ranges in panel C vs panel D. Quantitative data are presented as mean ± SD. Results are representative from 3 independent experiments (panels A-B), and the average of 3 independent experiments (panels C-D). Concentrations in brackets are in micromolar. ∗ P < .05; ∗∗ P < .01; ∗∗∗ P < .005; ∗∗∗∗ P < .0001. FC, FCCP; Glut, glutamate; LCL, LCL-805; ns, not significant; Oct-Carn, octanoyl-carnitine; SAC, SACLAC; Succ, succinate; VEN, venetoclax.

Article Snippet: HL-60 and MV4-11 cells were obtained from ATCC (Manassas, VA).

Techniques: Inhibition, Membrane, Flow Cytometry

Journal: Blood Neoplasia

Article Title: Acid ceramidase inhibition enhances BCL-2 targeting in venetoclax-resistant acute myeloid leukemia

doi: 10.1016/j.bneo.2026.100196

Figure Lengend Snippet: AC inhibition augmented the cytotoxicity of venetoclax and cytarabine. (A) MM-6 and MV4-11 VEN-R cells were treated with DMSO, SACLAC, venetoclax, cytarabine, doublet combinations, and triplet combinations at the indicated concentrations for 48 hours, then MTS cell viability was assessed. (B) Flow cytometry annexin V profiling of MM-6 and MV4-11 VEN-R cells treated with DMSO, SACLAC, venetoclax, cytarabine, doublet combinations, and triplet combinations at the indicated concentrations for 48 hours. Significance was assessed by Welch ANOVA with Dunnett multiple comparisons test. Quantitative data are presented as mean ± SD. Results are representative from 3 independent experiments. Concentrations in brackets are in micromolar. ∗ P < .05; ∗∗ P < .01; ∗∗∗ P < .005; ∗∗∗∗ P < .0001. AraC, cytarabine; ns, not significant; SAC, SACLAC; VEN, venetoclax.

Article Snippet: HL-60 and MV4-11 cells were obtained from ATCC (Manassas, VA).

Techniques: Inhibition, Flow Cytometry

Journal: Cell Reports Medicine

Article Title: Enalaprilat reverses neutrophil polarization imbalance via targeting taurine-STING axis for treatment of diabetic wounds

doi: 10.1016/j.xcrm.2026.102714

Figure Lengend Snippet: Regulatory roles of Ena in ZNF460-GGT1 axis-mediated taurine metabolism (A) KEGG pathway analysis of taurine metabolism. (B) Heatmap of top 40 significant DEGs. (C) Predicted binding sites of ZNF460 in the promoter of GGT1 . (D) Combination of ZNF460 with the promoter sequences of GGT1 as validated by ChIP assay. n = 3 independent experiments. (E) Dual luciferase reporter gene assay was conducted to evaluate the inhibitory effects of Ena on the conjunction of ZNF460 with GGT1 promoter. n = 3 independent experiments. (F) Molecular docking analysis between Ena and ZNF460. (G) Results of cellular thermal shift assay as visualized by western blot. n = 3 independent experiments. (H) Results of drug affinity responsive target stability test as visualized by western blot. n = 3 independent experiments. (I) Results of pull-down assay as visualized by western blot. n = 3 independent experiments. (J) Immunofluorescence staining was employed to assess the nuclear translocation of ZNF460 in HL-60 neutrophils with different treatments. Scale bar, 4 μm; n = 3 independent experiments. (K) Immunofluorescence staining was employed to assess the expression of GGT1 in HL-60 neutrophils with different treatments. Scale bar, 4 μm; n = 3 independent experiments. (L) After the cells underwent indicated treatments, GGT1 expression was detected. n = 3 independent experiments. (M) After the cells underwent indicated treatments, taurine content was detected. n = 3 independent experiments. (N) mRNA levels of cdo , csad , fmo1 , and baat as measured using RT-qPCR. n = 3 independent experiments. Data were shown as mean ± standard deviation (SD) from biological replicates, and statistical comparisons were performed using unpaired Student’s t test in (D, G, I, and N) and one-way ANOVA followed by Tukey’s multiple comparisons test in (D, E, H, and J–M). ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001, ∗∗∗∗ p < 0.0001.

Article Snippet: Neutrophil cell line (HL-60) and human umbilical vein endothelial cell (HUVEC) were gained from the American Type Culture Collection (ATCC) and grown at 37°C with 5% CO 2 .

Techniques: Binding Assay, Luciferase, Reporter Gene Assay, Thermal Shift Assay, Western Blot, Pull Down Assay, Immunofluorescence, Staining, Translocation Assay, Expressing, Quantitative RT-PCR, Standard Deviation

Journal: iScience

Article Title: A multimodal atlas for immunotherapeutic targeting of AML surface heterogeneity

doi: 10.1016/j.isci.2026.115337

Figure Lengend Snippet: Systematic ranking of antigens based on expression on AML blasts and non-hematopoietic tissue (A) Antigen intensity threshold for killing by surface targeting modalities. Left: Percent live cells of MOLM13 clones, indicated by their CD33 expression intensities, upon exposure to increasing concentrations of GO. Data are represented as mean ± SD, n = 4. Right: % dead cells following the incubation of MOLM13 clones with CD33-targeting CD33-bbz or CD33-28z CAR-T effector cells for 48 h. Data are represented as mean ± SD, n = 3. Dotted line indicates background target cell viability in the absence of effector cells. ∗∗∗ANOVA p < 0.001. (B) Heatmaps show estimated antigen count for the top 25 most highly expressed across 5,000 blast cells randomly sampled from diagnosis (left) and relapse (right) AML samples. Each column corresponds to an individual blast. Top annotation bar indicates the patient of origin. Higher antigen density for a specific antigen in a single blast is denoted by red shading. ∗Antibodies directed against CD13, CD45, CD47, CD99, and HLA-DR were found to be undersaturated (refer to “ ” in ). (C) Heatmaps showing gene expression of CD33, CLL-1, LAIR1, DEC-205, ITGA4, CD244, ADGRE2, and HER2 in non-hematopoietic cell types using single-cell RNA-seq data downloaded from Tabula Sapiens and GTEx databases. Columns are categorized based on cell types, and the top annotation bar indicates the tissue of origin of the cells. High and low relative expression are indicated by yellow and blue, respectively. Abbreviations: ANOVA, analysis of variance; GO, gene ontology; SD, standard deviation.

Article Snippet: Co-culture experiments of HDR CAR-T and MOLM13 clones and CAR-T and HL-60 (stable AML cell line, ATCC) target cells were conducted to investigate antigen-specific cytolysis at 1:1 E:T ratio.

Techniques: Expressing, Clone Assay, Incubation, Biomarker Discovery, Gene Expression, Single Cell, RNA Sequencing, Standard Deviation