human aml cell lines  (ATCC)

Journal: Scientific Reports

Article Title: The Caspase-1-EGR4 axis regulates macrophage repolarization in acute myeloid leukemia cells

doi: 10.1038/s41598-026-41381-x

Figure Lengend Snippet: Efficient CASP1 KD in THP-1 and MOLM-13 cells. ( A ) (left) GFP fluorescence and (right) FCM analysis of GFP expression levels in the stable polyclonal cell pools. ( B , C ) Evaluation of CASP1-KD efficiency by qPCR and western blot. Values represent mean ± SD of triplicate experiments. Statistical significance versus the shNC group is shown, with *, ***, and ns representing p < 0.05, p < 0.001, and not significant, respectively. Cropped blot images are shown. Groupings of lanes from non-adjacent parts of the same gel are indicated by black borders. Full-length, uncropped original blots are provided in Supplementary Dataset.

Article Snippet: The human AML cell lines THP-1 and MOLM-13 were purchased from Procell Life Science and Technology Co., Ltd (Wuhan, China).

Techniques: Fluorescence, Expressing, Western Blot

Journal: Scientific Reports

Article Title: The Caspase-1-EGR4 axis regulates macrophage repolarization in acute myeloid leukemia cells

doi: 10.1038/s41598-026-41381-x

Figure Lengend Snippet: Flow cytometric analysis of macrophage polarization markers. The THP-1 ( A , B ) and MOLM-13 ( C , D ) models are presented in each panel with representative histograms (left) and quantitative data (right). ( A , C ) CD163 + M2 macrophages; ( B , D ) CD86 + M1 macrophages. Values shown are mean ± SD ( n ≥ 3 independent experiments). *, **, ***, or ns denotes p < 0.05, p < 0.01, p < 0.001,or not significant versus the shNC control, respectively.

Article Snippet: The human AML cell lines THP-1 and MOLM-13 were purchased from Procell Life Science and Technology Co., Ltd (Wuhan, China).

Techniques: Control

Journal: Scientific Reports

Article Title: The Caspase-1-EGR4 axis regulates macrophage repolarization in acute myeloid leukemia cells

doi: 10.1038/s41598-026-41381-x

Figure Lengend Snippet: Transcriptomic profiling identifies EGR4 as a key transcriptional target upregulated by CASP1 KD in AML cells. ( A ) Heat plot of the global transcriptome from shNC and shCASP1 THP-1 cells. ( B ) Distribution of DEGs is shown in a volcano plot, where statistically significant upregulations and downregulations are highlighted in red and blue, respectively. ( C ) KEGG pathway (Kanehisa Laboratories, https://www.kegg.jp/ ) classification of the DEGs, showing the most enriched functional categories. ( D ) GO (The Gene Ontology Resource, http://geneontology.org/ ) enrichment analysis for biological processes, showing the most significantly enriched terms. ( E ) qPCR validation of EGR4 mRNA expression in THP-1 cells from the indicated groups. ( F , G ) EGR4 protein expression was assessed by western blot in THP-1 and MOLM-13 cells. We calculated the data from three independent experiments as the mean ± SD, using *** p < 0.001 as the threshold for statistical significance compared to the shNC group. Cropped blot images are shown. Groupings of lanes from non-adjacent parts of the same gel are indicated by black borders. Full-length, uncropped original blots are provided in Supplementary Dataset.

Article Snippet: The human AML cell lines THP-1 and MOLM-13 were purchased from Procell Life Science and Technology Co., Ltd (Wuhan, China).

Techniques: Functional Assay, Biomarker Discovery, Expressing, Western Blot

Journal: Scientific Reports

Article Title: The Caspase-1-EGR4 axis regulates macrophage repolarization in acute myeloid leukemia cells

doi: 10.1038/s41598-026-41381-x

Figure Lengend Snippet: EGR4 KD rescues the macrophage polarization shift induced by CASP1 KD. ( A , C ) RT-qPCR analysis of EGR4 mRNA levels. ( B , D ) Representative immunoblot of EGR4 protein expression. ( E , F ) Representative immunoblots for CD206 and CD86 in macrophages following stimulation with CM from the indicated AML cell groups. ( G ) Representative immunofluorescence staining for CD206 (red) in macrophages.Nuclear staining was performed with DAPI (blue) (scale bar: 50 μm). ( H – K ) Flow cytometric analysis of ( H , J ) CD163 + and ( I , K ) CD86 + macrophage populations after treatment with CM from ( H , I ) THP-1 or ( J , K ) MOLM-13 models. Graphical summary shows representative histograms (left) and the corresponding quantification of positive cells (right), presented as mean ± SD ( n ≥ 3). Cropped blot images are shown. Groupings of lanes from non-adjacent parts of the same gel are indicated by black borders. Full-length, uncropped original blots are provided in Supplementary Dataset.

Article Snippet: The human AML cell lines THP-1 and MOLM-13 were purchased from Procell Life Science and Technology Co., Ltd (Wuhan, China).

Techniques: Quantitative RT-PCR, Western Blot, Expressing, Immunofluorescence, Staining

Journal: Scientific Reports

Article Title: The Caspase-1-EGR4 axis regulates macrophage repolarization in acute myeloid leukemia cells

doi: 10.1038/s41598-026-41381-x

Figure Lengend Snippet: Mechanism of the CASP1-EGR4 axis through the IL-10/STAT3 pathway in macrophages. ( A , B ) Conditioned medium (CM) from THP-1 cells with CASP1 knockdown (KD) reduces the levels of IL-10 and phosphorylated STAT3 (p-STAT3), but not total STAT3, in recipient macrophages. ( A ) Representative western blots and ( B ) statistical summary. ( C , D ) CM from MOLM-13 cells with CASP1 KD reduces the levels of IL-10 and p-STAT3, but not total STAT3, in recipient macrophages. ( C ) Representative western blots and ( D ) statistical summary. ( E , F ) Additional KD of EGR4 in CASP1-KD THP-1 cells partially reverses the reduction of IL-10 and p-STAT3 in macrophages treated with the corresponding CM. ( E ) Representative western blots and ( F ) statistical summary. ( G , H ) Additional KD of EGR4 in CASP1-KD MOLM-13 cells partially reverses the reduction of IL-10 and p-STAT3 in macrophages treated with the corresponding CM. ( G ) Representative western blots and ( H ) statistical summary. Cropped blot images are shown. The STAT3 and p-STAT3 data are presented in separate frames to indicate they are from different blots. Full-length, uncropped original blots and, where informative, multiple exposures (e.g., for IL-10 and STAT3) are provided in Supplementary Dataset.

Article Snippet: The human AML cell lines THP-1 and MOLM-13 were purchased from Procell Life Science and Technology Co., Ltd (Wuhan, China).

Techniques: Knockdown, Western Blot

Journal: Journal of Cell Communication and Signaling

Article Title: Mechanistic role of GNE‐987 targeting BRD4‐HCP5 axis in pediatric T‐cell acute lymphoblastic leukemia

doi: 10.1002/ccs3.70063

Figure Lengend Snippet: BRD4 inhibitor significantly promotes HCP5 super‐enhancer activity and expression. (A) Reverse transcription‐quantitative polymerase chain reaction analysis of HCP5 mRNA expression levels in acute myeloid leukemia cell lines (THP‐1 and HL‐60) treated with the BRD4 inhibitor GNE‐987. (B) Western Blot analysis showing BRD4 expression levels in different groups. (C) Representative agarose gel electrophoresis image and statistical quantification of ChIP‐qPCR products. Compared to the Ctrl group, * p < 0.05, ** p < 0.01. All cell experiments were repeated three times. BRD4, Bromodomain‐containing protein 4; HCP5, HLA Complex P5.

Article Snippet: The acute myeloid leukemia (AML) cell lines Tsuchiya Human Phagocyte‐1 (THP‐1) and Human Leukemia‐60 (HL‐60) (TIB‐202 and CCL‐240, ATCC), along with T‐ALL–specific cell lines Jurkat, CCRF‐CEM, MOLT‐4, and RPMI‐8402 (TIB‐152, CCL‐111, CRL‐1582, and CCL‐27, ATCC), were cultured in RPMI‐1640 medium (11875093, Thermo Fisher Scientific) supplemented with 10% fetal bovine serum (A5670701, Gibco) and maintained in an incubator at 37°C with 5% CO 2 .

Techniques: Activity Assay, Expressing, Reverse Transcription, Real-time Polymerase Chain Reaction, Western Blot, Agarose Gel Electrophoresis, ChIP-qPCR

Journal: Journal of Cell Communication and Signaling

Article Title: Mechanistic role of GNE‐987 targeting BRD4‐HCP5 axis in pediatric T‐cell acute lymphoblastic leukemia

doi: 10.1002/ccs3.70063

Figure Lengend Snippet: Bromodomain‐containing protein 4 Inhibitor GNE‐987 Significantly Inhibits acute myeloid leukemia Cell Proliferation and Migration and Induces Apoptosis. (A) CCK‐8 assay measuring cell proliferation in each group at 0, 12, 24, 36, 48, 60, and 72 h, with absorbance detected at OD450. (B) Representative images of Live and Dead staining for each group, with a bar chart depicting the statistical analysis of cell death ratios. Bar = 50 μm. (C) Colony formation assay for each group, with a bar chart showing the statistical analysis of colony numbers. (D) Flow cytometry analysis of apoptosis levels in each group, with a bar chart depicting the statistical analysis of apoptosis rates. Compared with the Ctrl group, * p < 0.05, ** p < 0.01. All cell experiments were repeated three times.

Article Snippet: The acute myeloid leukemia (AML) cell lines Tsuchiya Human Phagocyte‐1 (THP‐1) and Human Leukemia‐60 (HL‐60) (TIB‐202 and CCL‐240, ATCC), along with T‐ALL–specific cell lines Jurkat, CCRF‐CEM, MOLT‐4, and RPMI‐8402 (TIB‐152, CCL‐111, CRL‐1582, and CCL‐27, ATCC), were cultured in RPMI‐1640 medium (11875093, Thermo Fisher Scientific) supplemented with 10% fetal bovine serum (A5670701, Gibco) and maintained in an incubator at 37°C with 5% CO 2 .

Techniques: Migration, CCK-8 Assay, Staining, Colony Assay, Flow Cytometry

Journal: Journal of Cell Communication and Signaling

Article Title: Mechanistic role of GNE‐987 targeting BRD4‐HCP5 axis in pediatric T‐cell acute lymphoblastic leukemia

doi: 10.1002/ccs3.70063

Figure Lengend Snippet: Silencing of HLA Complex P5 Reverses the Inhibitory Effect of GNE‐987 on acute myeloid leukemia Cell Viability. (A) Reverse transcription‐quantitative polymerase chain reaction results verify the silencing efficiency of sh‐HCP5. (B) CCK8 assay measuring cell proliferation at 0, 12, 24, 36, 48, 60, and 72 h, with absorbance detected at OD450. (C) Representative images of Live and Dead staining in each group and a bar chart showing the death ratio; scale bar = 50 μm. (D) Colony formation assay and statistical graph of colony numbers for each group. (E) Flow cytometry analysis of apoptosis levels and a statistical graph of apoptosis rates in each group. For panel A, compared with the sh‐NC group, * p < 0.05, ** p < 0.01. For panels B‐E, compared with the Ctrl group, * p < 0.05, ** p < 0.01; compared with the GNE‐987+sh‐NC group, # p < 0.05, ## p < 0.01. All cell experiments were repeated three times.

Article Snippet: The acute myeloid leukemia (AML) cell lines Tsuchiya Human Phagocyte‐1 (THP‐1) and Human Leukemia‐60 (HL‐60) (TIB‐202 and CCL‐240, ATCC), along with T‐ALL–specific cell lines Jurkat, CCRF‐CEM, MOLT‐4, and RPMI‐8402 (TIB‐152, CCL‐111, CRL‐1582, and CCL‐27, ATCC), were cultured in RPMI‐1640 medium (11875093, Thermo Fisher Scientific) supplemented with 10% fetal bovine serum (A5670701, Gibco) and maintained in an incubator at 37°C with 5% CO 2 .

Techniques: Reverse Transcription, Real-time Polymerase Chain Reaction, CCK-8 Assay, Staining, Colony Assay, Flow Cytometry

Journal: EMBO Molecular Medicine

Article Title: Dual targeting of CDK6 and LSD1 is synergistic and overcomes differentiation blockade in AML

doi: 10.1038/s44321-025-00296-2

Figure Lengend Snippet: ( A ) Experimental design for the treatment of AML cells. AML cells, cocultured with HS-5 stromal cells, were treated for 4 days with palbociclib at 0.5 μM and/or TCP at 5 μM, and analyzed by flow cytometry and May-Grünwald–Giemsa staining. ( B ) The percentage of MV4-11 cells expressing CD11b ( n = 3), CD14 ( n = 3) or CD86 ( n = 4) myeloid markers was quantified by flow cytometry following the indicated treatments. Ctrl control, TCP tranylcypromine, Palbo palbociclib, Combo is the combination of palbociclib with tranylcypromine. The exact adjusted p values were as follows: Left panel, Ctrl vs Combo p < 0.0001, TCP vs Combo p = 0.0001, Palbo vs Combo p = 0.0001. Middle panel: Ctrl vs Combo p = 0.0001, TCP vs Combo p = 0.0001, Palbo vs Combo p = 0.0008. Right panel: Ctrl vs Combo p = 0.0002, TCP vs Combo p = 0.0003, Palbo vs Combo p = 0.0023. ( C – E ) Expression of the late monocytic marker CD86 on MOLM-14 ( C ) ( n = 3), PL-21 ( D ) ( n = 3), and THP-1 ( E ) ( n = 3) AML cell lines. The percentage of cells expressing CD86 was quantified by flow cytometry following the indicated treatments. The exact adjusted p values were as follows: ( C ) Ctrl vs Combo p = 0.0003, TCP vs Combo p = 0.0005, Palbo vs Combo p = 0.0019. ( D ) Ctrl vs Combo p < 0.0001, TCP vs Combo p < 0.0001, Palbo vs Combo p = 0.0117, Ctrl vs Palbo p = 0.0005, TCP vs Palbo p = 0.0012. ( E ) Ctrl vs Combo p = 0.0005, TCP vs Combo p = 0.0019, Palbo vs Combo p = 0.0098, Ctrl vs Palbo p = 0.0297. ( F ) Cell morphology of the AML cell lines following the treatments. AML cell lines were stained with May-Grünwald–Giemsa. Cells were observed under a Zeiss Apotome microscope. The indicated scale is 50 µm. ( G ) Primary AML patient cells were treated with the indicated treatments for 96 h. After treatment, cells were analyzed by flow cytometry for the expression of CD11b ( n = 17) and CD86 ( n = 14). Each colored dot represents a sample. Exact adjusted pvalues: left panel, Ctrl vs TCP p = 0.0070, Ctrl vs Palbo p = 0.0059, Ctrl vs Combo p < 0.0001, TCP vs Combo p = 0.0004, Palbo vs Combo p = 0.0005; right panel, Ctrl vs Combo p < 0.0001, TCP vs Combo p = 0.0001, Palbo vs Combo p = 0.0022. ( H ) May-Grünwald–Giemsa staining of five primary AML patient samples following the indicated treatments. The indicated scale is 50 µm. Histograms indicate the mean of independent experiments ± SD. Statistical analyses were performed using a one-way ANOVA and followed by Tukey’s test. * p < 0.05, ** p < 0.01, *** p < 0.001, and **** p < 0.0001.

Article Snippet: The human AML cell lines PL-21 (ACC 536) and SKM1 (ACC 547) were obtained from DSMZ, and the human AML cell line THP-1 (TIB-202) was purchased from ATCC.

Techniques: Flow Cytometry, Staining, Expressing, Control, Marker, Microscopy

Journal: EMBO Molecular Medicine

Article Title: Dual targeting of CDK6 and LSD1 is synergistic and overcomes differentiation blockade in AML

doi: 10.1038/s44321-025-00296-2

Figure Lengend Snippet: ( A ) CFSE cell proliferation analysis in PL-21 AML cell line. Cells treated as described before for 96 h, were labeled with CFSE to monitor cell division over the following 6 days. ( B ) Serial plating assays of MV4-11 on methylcellulose following a single round of treatment. MV4-11 cells were treated as previously. After the 4-day treatments, an equal number of cells was seeded in methylcellulose for 10 days ( n = 3), and two serial replatings were performed (second and third plating; n = 3 and n = 2, respectively) without any additional treatment. Left panel, Ctrl vs Palbo p = 0.0024, Ctrl vs Combo p < 0.0001, TCP vs Palbo p = 0.0303, TCP vs Combo p = 0.0002, Palbo vs Combo p = 0.0035. Middle panel, Ctrl vs Combo p = 0.0028, TCP vs Combo p = 0.0032, Palbo vs Combo p = 0.0104. ( C ) Serial treatments and replating assays of MV4-11 cells. Top panel: Experimental design for the treatments and replating strategy. After the 4-day treatment cycle, MV4-11 cells were seeded into methylcellulose for 10 days. Bottom panel: quantification of the remaining colonies over three rounds of replating. First plating, n = 3; second plating, n = 3; third plating, n = 2. Left panel, Ctrl vs Palbo p = 0.0147, Ctrl vs Combo p = 0.0005, TCP vs Combo p = 0.0037, Palbo vs Combo p = 0.0229. Middle panel, Ctrl vs Palbo p = 0.0005, Ctrl vs Combo p = 0.0001, TCP vs Palbo p = 0.0024, TCP vs Combo p = 0.0004. ( D ) Representative pictures of colonies obtained in the third round of replating of MV4-11 cells (top panel) and MOLM-14 cells (bottom panel). Statistical analyses are performed using a one-way ANOVA test followed by Tukey’s test. * p < 0.05, ** p < 0.01, *** p < 0.001, and **** p < 0.0001.

Article Snippet: The human AML cell lines PL-21 (ACC 536) and SKM1 (ACC 547) were obtained from DSMZ, and the human AML cell line THP-1 (TIB-202) was purchased from ATCC.

Techniques: Labeling