acute monocytic leukemia cell line thp 1  (DSMZ)

Journal: Regenerative Therapy

Article Title: WTAP stabilizes MMP12 expression to promote the malignant phenotypes of esophageal cancer cells

doi: 10.1016/j.reth.2026.101101

Figure Lengend Snippet: MMP12 silencing inhibited M2 macrophage polarization. THP-1 cells were differentiated into M0 macrophages (THP-1 M0) by treatment with 100 ng/mL PMA for 24 h. (A) Flow cytometry was used to quantify the number of CD68-positive cells. Subsequently, KYSE150 cells were co-cultured with the THP-1-derived macrophages using a Transwell system. (B) The mRNA levels of IL-10, Arg-1, and TGF-β were detected by qRT-PCR. (C) Flow cytometry was used to quantify the number of CD206-positive macrophages. (D) Cell migration analysis by transwell migration assay. ∗ P < 0.05, ∗∗ P < 0.01 and ∗∗∗ P < 0.001.

Article Snippet: For immunophenotyping, single-cell suspensions of both THP-1 and THP-1-M0 cells were labeled with an anti-CD68 antibody (E-AB-F1299L, Elabscience, Wuhan, China).

Techniques: Flow Cytometry, Cell Culture, Derivative Assay, Quantitative RT-PCR, Migration, Transwell Migration Assay

Journal: Regenerative Therapy

Article Title: WTAP stabilizes MMP12 expression to promote the malignant phenotypes of esophageal cancer cells

doi: 10.1016/j.reth.2026.101101

Figure Lengend Snippet: WTAP silencing inhibited M2 macrophage polarization by regulating MMP12. THP-1 cells were differentiated into M0 macrophages (THP-1 M0) by treatment with 100 ng/mL PMA for 24 h. KYSE150 cells were transfected with si-WTAP, MMP12 overexpression plasmid, or the matched control (si-NC and oe-NC). Subsequently, these KYSE150 cells were co-cultured with the THP-1-derived macrophages using a Transwell system. (A) The mRNA levels of IL-10, Arg-1, and TGF-β were detected by qRT-PCR. (B) Flow cytometry was used to quantify the number of CD206-positive macrophages. (C) Cell migration analysis by transwell migration assay. ∗ P < 0.05, ∗∗ P < 0.01 and ∗∗∗ P < 0.001.

Article Snippet: For immunophenotyping, single-cell suspensions of both THP-1 and THP-1-M0 cells were labeled with an anti-CD68 antibody (E-AB-F1299L, Elabscience, Wuhan, China).

Techniques: Transfection, Over Expression, Plasmid Preparation, Control, Cell Culture, Derivative Assay, Quantitative RT-PCR, Flow Cytometry, Migration, Transwell Migration Assay

Journal: Translational Oncology

Article Title: REST-driven upregulation of SFXN3 promotes AML progression via Wnt/β-catenin activation and confers decitabine resistance

doi: 10.1016/j.tranon.2026.102705

Figure Lengend Snippet: Effects of SFXN3 Knockdown on Proliferation, Apoptosis, and Signaling Pathways in AML Cells. (A–C) qRT-PCR and Western blot analyses were used to measure SFXN3 expression levels in various leukemia cell lines (THP-1, KG-1, U937, K562) and in normal bone marrow stromal cells (HS-5). (D) Two independent shRNAs (sh-SFXN3–1 and sh-SFXN3–2) were used to knock down SFXN3 expression in THP-1 and KG-1 cells. Western blot was performed to assess the knockdown efficiency and specificity. (E) Quantification of SFXN3 knockdown efficiency by different shRNAs. (F) CCK-8 cell proliferation assays were conducted to evaluate the effects of SFXN3 knockdown on cell growth dynamics over time. (G) EdU incorporation assays were used to assess DNA synthesis activity, indirectly reflecting cellular proliferation, and to compare differences between knockdown and control groups, (bar=50ųm). (H) Western blot analysis of key cell cycle regulatory proteins (CDK4, CDK6, P27, and P21) to investigate the potential mechanism by which SFXN3 affects cell cycle progression. (I) TUNEL assays were used to evaluate apoptosis levels in the knockdown versus control groups, assessing the role of SFXN3 in apoptosis suppression, (bar=50ųm). (J) Western blot analysis of pro-apoptotic proteins (BAX and BAK) and anti-apoptotic proteins (Bcl-2 and Bcl-xl) in THP-1 and KG-1 cells following SFXN3 knockdown. (K) Correlation analysis between SFXN3 expression and key proteins in the Wnt/β-Catenin signaling pathway. (L) Subcellular fractionation followed by Western blotting was performed to assess β-catenin nuclear translocation. Cytoplasmic (Cyto) and nuclear (Nuc) fractions were probed for β-catenin, with β-actin (cytoplasmic marker) and Histon H3 (nuclear marker) used to confirm fractionation quality. Data are presented as mean ± SD. from three independent experiments ( n = 3). One-way ANOVA was used in (A, B, E), and two-way ANOVA was used in (F). *, p < 0.05.

Article Snippet: The ATCC supplied the AML cell lines THP-1, KG-1, U937, and K562, and the stromal cell line HS-5.

Techniques: Knockdown, Protein-Protein interactions, Quantitative RT-PCR, Western Blot, Expressing, CCK-8 Assay, DNA Synthesis, Activity Assay, Control, TUNEL Assay, Fractionation, Translocation Assay, Marker

Journal: Translational Oncology

Article Title: REST-driven upregulation of SFXN3 promotes AML progression via Wnt/β-catenin activation and confers decitabine resistance

doi: 10.1016/j.tranon.2026.102705

Figure Lengend Snippet: The Wnt/β-Catenin Pathway Agonist SKL2001 Reverses the Effects of SFXN3 Knockdown on Leukemia Cell Proliferation and Apoptosis. (A) Western blot analysis of SFXN3 protein expression following SFXN3 knockdown and treatment with SKL2001, to assess whether SKL2001 significantly modulates SFXN3 expression. (B) CCK-8 assays were performed to evaluate whether SKL2001 could reverse the inhibitory effects of SFXN3 knockdown on the proliferation of THP-1 and KG-1 leukemia cells. (C) EdU staining assays were used to assess DNA synthesis activity, analyzing the ability of SKL2001 to restore proliferation suppressed by SFXN3 knockdown, (bar=50 ųm). (D) Quantitative analysis of EdU fluorescence intensity to evaluate DNA replication across different treatment groups. (E) Western blot analysis of cell cycle regulators CDK4, CDK6, Cyclin D1, and Cyclin E1 to determine whether SKL2001 rescues the expression of these proteins in SFXN3-silenced cells. (F) Western blot analysis of pro-apoptotic proteins (BAX, BAK) and anti-apoptotic proteins (Bcl-2, Bcl-xl) to confirm that SKL2001 mitigates the apoptosis-promoting effects of SFXN3 knockdown. (G) TUNEL assays were conducted to assess whether SKL2001 suppresses the enhanced apoptosis induced by SFXN3 knockdown, (bar=50ųm). (H) Quantification of TUNEL fluorescence intensity, reflecting apoptosis levels under different treatment conditions. (I) Subcellular fractionation followed by Western blotting was performed to assess β-catenin nuclear translocation. Cytoplasmic (Cyto) and nuclear (Nuc) fractions were probed for β-catenin, with β-actin (cytoplasmic marker) and Histon H3 (nuclear marker) used to confirm fractionation quality. Data are presented as mean ± SD. from at least three independent experiments. One-way ANOVA was used in (D, H), and two-way ANOVA was used in (B). *, p < 0.05; **, p < 0.01; ***, p < 0.001 vs. control or scramble group.

Article Snippet: The ATCC supplied the AML cell lines THP-1, KG-1, U937, and K562, and the stromal cell line HS-5.

Techniques: Knockdown, Western Blot, Expressing, CCK-8 Assay, Staining, DNA Synthesis, Activity Assay, Fluorescence, TUNEL Assay, Fractionation, Translocation Assay, Marker, Control

Journal: Translational Oncology

Article Title: REST-driven upregulation of SFXN3 promotes AML progression via Wnt/β-catenin activation and confers decitabine resistance

doi: 10.1016/j.tranon.2026.102705

Figure Lengend Snippet: The REST–SFXN3 Axis Promotes Malignant Phenotypes in AML Cells via the Wnt/β-Catenin Signaling Pathway. (A) Western blot analysis of the effect of REST knockdown (sh-REST) on SFXN3 expression, and the reversal of this effect by SFXN3 overexpression. (B) CCK-8 assays assess the impact of sh-REST and SFXN3 overexpression on AML cell proliferation. (C) EdU incorporation assays evaluate the effects of sh-REST and SFXN3 overexpression on DNA synthesis activity in AML cells, (bar=50ųm). (D) Quantification of EdU-positive cells to compare DNA synthesis capacity across groups. (E) Western blot analysis of proliferation-related proteins CDK4, CDK6, Cyclin D1, and Cyclin E1 under sh-REST and SFXN3 overexpression conditions. (F) Band intensities were quantified using ImageJ software and normalized to the indicated internal controls. (G) TUNEL assays detect apoptotic cells after REST knockdown and SFXN3 overexpression, (bar=50ųm). (G) Quantitative analysis of apoptotic cells in THP-1 and KG-1 cell lines. (H) Quantification of TUNEL fluorescence intensity, reflecting apoptosis levels under different treatment conditions. (I) Western blot evaluation of pro-apoptotic proteins (BAX, BAK) and anti-apoptotic proteins (Bcl-2, Bcl-xl), demonstrating REST knockdown promotes apoptosis, which is reversed by SFXN3 overexpression. (J) Subcellular fractionation followed by Western blotting was performed to assess β-catenin nuclear translocation. Cytoplasmic (Cyto) and nuclear (Nuc) fractions were probed for β-catenin, with β-actin (cytoplasmic marker) and Histon H3 (nuclear marker) used to confirm fractionation quality. Data are presented as mean ± SD from three independent experiments ( n = 3).One-way ANOVA was used in (D, F,H), and two-way ANOVA was used in (B). **, p < 0.01; ***, p < 0.001.

Article Snippet: The ATCC supplied the AML cell lines THP-1, KG-1, U937, and K562, and the stromal cell line HS-5.

Techniques: Western Blot, Knockdown, Expressing, Over Expression, CCK-8 Assay, DNA Synthesis, Activity Assay, Software, TUNEL Assay, Fluorescence, Fractionation, Translocation Assay, Marker

Journal: Translational Oncology

Article Title: REST-driven upregulation of SFXN3 promotes AML progression via Wnt/β-catenin activation and confers decitabine resistance

doi: 10.1016/j.tranon.2026.102705

Figure Lengend Snippet: Decitabine Suppresses AML Cell Proliferation and Promotes Apoptosis via SFXN3 Inhibition. (A) RT-PCR analysis of the effects of Gefitinib, Disulfiram, and Decitabine on SFXN3 mRNA expression. (B) Western blot analysis of SFXN3 protein levels following treatment with Gefitinib, Disulfiram, and Decitabine. (C) CCK-8 assay to calculate the IC50 values of Decitabine in THP-1 and KG-1 cells, identifying appropriate drug concentrations for subsequent experiments (D) CCK-8 assays were performed to evaluate AML cell viability at 6,12,24,48, and 72 h following treatment with 50 nm decitabine, thereby determining the optimal treatment duration. E) EdU incorporation assay evaluating the proliferation capacity of AML cells after Decitabine treatment, (bar=50ųm). (F) Western blot analysis of proliferation-related proteins (P21, P27, CDK4, and CDK6) following Decitabine treatment. (G) TUNEL staining to detect DNA fragmentation at the 3′-OH ends, marking apoptotic cells after Decitabine exposure, (bar=50ųm). (H) Western blot analysis of pro-apoptotic (e.g., BAX, BAK) and anti-apoptotic (e.g., Bcl-2, Bcl-xl) protein expression in response to Decitabine. (I) Western blot analysis of key components of the Wnt/β-Catenin signaling pathway after Decitabine treatment, revealing pathway inhibition. Subcellular fractionation followed by Western blotting was performed to assess β-catenin nuclear translocation. Cytoplasmic (Cyto) and nuclear (Nuc) fractions were probed for β-catenin, with β-actin (cytoplasmic marker) and Histon H3 (nuclear marker) used to confirm fractionation quality. n = 3,Error bars indicate mean ± SD; One-way ANOVA in (D, F); **, p < 0.01, *** p <0.001.

Article Snippet: The ATCC supplied the AML cell lines THP-1, KG-1, U937, and K562, and the stromal cell line HS-5.

Techniques: Inhibition, Reverse Transcription Polymerase Chain Reaction, Expressing, Western Blot, CCK-8 Assay, TUNEL Assay, Staining, Fractionation, Translocation Assay, Marker

Journal: Cell Insight

Article Title: TNF inhibits SARS-CoV-2 induced cell-cell fusion through activating the SDC4-RhoA signaling to promote actin bundles formation

doi: 10.1016/j.cellin.2026.100310

Figure Lengend Snippet: Host factors secreted by innate immune cells early after activation inhibit SARS-CoV-2 spike protein induced cell-cell fusion. (A) Luciferase assay showing the effect of supernatant from THP-1 cell cultures pretreated with the NLRP3 inhibitor MCC950 (10 μM) and then stimulated with the TLR1/2 ligand Pam3CSK4 (1 μg/mL) for 3 h on spike protein-induced HEK293T cell-cell fusion. Serum-free RPMI 1640 served as the medium control. Data points represent mean ± SEM from six independent experiments; P values are indicated. (B) Immunoblot analysis of S2′ cleavage in fused cells treated with supernatant from MCC950-pretreated THP-1 cultures stimulated with Pam3CSK4 for 3 h. Blots are representative of three independent experiments. (C) Luciferase assay showing the effect of supernatant from NLRP3-knockout THP-1 cell cultures stimulated with Pam3CSK4 for 3 h on spike-induced HEK293T cell-cell fusion. Serum-free RPMI 1640 served as the control. Data points represent mean ± SEM from six independent experiments; P values are indicated. (D) Immunoblot analysis of S2′ cleavage in fused cells treated with supernatant from NLRP3-knockout THP-1 cell cultures stimulated with Pam3CSK4 for 3 h. Blots are representative of three independent experiments. (E) Visualization of syncytium formation by ZsGreen fluorescence after treatment with supernatant from MCC950-pretreated THP-1 cultures stimulated with Pam3CSK4 for 3 h. Images are representative of three independent experiments; white arrows indicate syncytia. Scale bar, 50 μm. (F) Visualization of syncytium formation by ZsGreen fluorescence after treatment with supernatant from NLRP3-knockout THP-1 cell cultures stimulated with Pam3CSK4 for 3 h. Images are representative of three independent experiments; white arrows indicate syncytia. Scale bar, 50 μm. (G) Quantification of relative syncytium area (%) based on ZsGreen fluorescence images in (E). Data are presented as mean ± SEM from three independent experiments. P values are indicated above the bars. (H) Quantification of relative syncytium area (%) based on ZsGreen fluorescence images in (F). Data are presented as mean ± SEM from three independent experiments. P values are indicated.

Article Snippet: The human monocytic THP-1 cell line (TIB-202; ATCC) was authenticated via short tandem repeat (STR) analysis by Suzhou Genetic Testing Biotech Co., Ltd, following the ANSI/ATCC ASN-0002-2012 standard ( ; ).

Techniques: Activation Assay, Luciferase, Control, Western Blot, Knock-Out, Fluorescence

Journal: Cell Insight

Article Title: TNF inhibits SARS-CoV-2 induced cell-cell fusion through activating the SDC4-RhoA signaling to promote actin bundles formation

doi: 10.1016/j.cellin.2026.100310

Figure Lengend Snippet: TNF produced by innate immune cells early after activation inhibit SARS-CoV-2 spike induced cell-cell fusion. (A) Luciferase assay showing the effect of recombinant IL-6 (10 ng/mL), IL-8 (10 ng/mL), or TNF (10 ng/mL) on spike-induced cell-cell fusion. PBS was used as the vehicle control. Data points represent mean ± SEM from four independent experiments; P values are indicated. (B) Luciferase assay showing the effect of supernatant from THP-1 cultures stimulated with Pam3CSK4 for the indicated durations on cell-cell fusion in HEK293T cells pretreated with the IL-1 receptor antagonist (IL-1RA) (4 μg/mL). Data represent mean ± SEM from four independent experiments; P values are shown. (C) Immunoblot analysis of S2′ cleavage in IL-1RA-pretreated fused cells exposed to supernatant from THP-1 cultures stimulated with Pam3CSK4 for the indicated durations. Blots are representative of three independent experiments. (D) ELISA quantification of IL-1β and TNF levels in supernatant from THP-1 cell cultures after stimulation with the indicated TLR ligand at the specified time points. Data represent mean ± SEM from three independent experiments. (E) Luciferase assay showing the effect of supernatant from THP-1 cultures stimulated with Pam3CSK4 for the indicated durations on spike-induced fusion in HEK293T-sgcontrol and HEK293T-sgTNFR1 cells. Data points represent mean ± SEM from six independent experiments; P values are indicated. (F) Immunoblot analysis of S2′ cleavage in HEK293T-sgcontrol and HEK293T-sgTNFR1 fused cells treated with supernatant from THP-1 cell cultures stimulated with Pam3CSK4 for the indicated durations. Blots are representative of three independent experiments. (G) Luciferase assay showing the effect of supernatant from Pam3CSK4-stimulated THP-1 cell cultures on cell-cell fusion in IL-1RA-pretreated HEK293T-sgcontrol and HEK293T-sgTNFR1 cells. Data represent mean ± SEM from four independent experiments; P values are indicated. (H) Immunoblot analysis of S2′ cleavage in IL-1RA-pretreated HEK293T-sgcontrol and HEK293T-sgTNFR1 fused cells exposed to supernatant from Pam3CSK4-stimulated THP-1 cell cultures. Blots are representative of three independent experiments. (I–J) ELISA quantification of IL-1β and TNF levels in supernatant from THP-1 cell cultures under (I) MCC950 pharmacological inhibition of NLRP3, (J) NLRP3 knockout, after stimulation with the indicated TLR ligand at the specified time points. Data represent mean ± SEM from three independent experiments.

Article Snippet: The human monocytic THP-1 cell line (TIB-202; ATCC) was authenticated via short tandem repeat (STR) analysis by Suzhou Genetic Testing Biotech Co., Ltd, following the ANSI/ATCC ASN-0002-2012 standard ( ; ).

Techniques: Produced, Activation Assay, Luciferase, Recombinant, Control, Western Blot, Enzyme-linked Immunosorbent Assay, Inhibition, Knock-Out