human monocytic leukemia cell line  (ATCC)

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: Regenerative Therapy

Article Title: Lung cancer cell-derived exosomal EHF drives M2 macrophage polarization via transcriptional activation of RNF41 to promote tumor progression

doi: 10.1016/j.reth.2026.101104

Figure Lengend Snippet: EHF promotes lung cancer cell malignancy and M2 macrophage polarization. A549 and H520 cells were transfected with control (Ctrl) or EHF-knockdown (KD-EHF) plasmids. (A-B) Western blot and quantification of EHF protein levels in A549 and H520 cells. (C-D) EdU staining and quantification of EdU + cells in A549 and H520 cells, assessing proliferation. (E-H) Glucose uptake and lactate production assays were conducted using commercial kits to evaluate glycolytic output. (I-J) Flow cytometric analysis of apoptosis. (K-L) Transwell assay was employed to examine cell migration. (M − P) THP-1 cells were differentiated into THP-1-M0 macrophages with 100 ng/mL PMA for 24 h. Culture supernatants from Ctrl- or KD-EHF–transfected A549 and H520 cells were collected and co-incubated with THP-1-M0 macrophages. (M − N) Flow cytometric analysis was performed to assess the proportion of CD206 + M2-polarized macrophages. (O–P) ELISA was conducted to quantify the secretion of M2-associated cytokines (TGF-β1, IL-10, and VEGFA) in the supernatants of THP-1-M0 macrophages. Data are presented as mean ± SD. P < 0.05 denotes statistical significance.

Article Snippet: Human monocyte cell line (THP-1) (Catalog #: YC-D011) were purchased from Guangzhou Ubigene Biosciences Co., Ltd. (Guangzhou, China) and cultured in RPMI-1640 (Gibco) containing 10% FBS (Gibco).

Techniques: Transfection, Control, Knockdown, Western Blot, Staining, Transwell Assay, Migration, Incubation, Enzyme-linked Immunosorbent Assay

Journal: Regenerative Therapy

Article Title: Lung cancer cell-derived exosomal EHF drives M2 macrophage polarization via transcriptional activation of RNF41 to promote tumor progression

doi: 10.1016/j.reth.2026.101104

Figure Lengend Snippet: Exosomes derived from lung cancer cells are internalized by THP-1-M0 macrophages and mediate EHF delivery. Exosomes were isolated from A549 and H520 cells. (A) TEM images showed the typical cup-shaped morphology of exosomes isolated from A549 and H520 cell supernatants (scale bar: 100 nm). (B) NTA of exosomes from A549 and H520 cells. (C) Western blot analysis of exosome markers (CD9 and TSG101) and the endoplasmic reticulum marker Calnexin in exosome lysates and parental A549/H520 cell lysates. (D-E) THP-1-M0 macrophages (differentiated from THP-1 cells with 100 ng/mL PMA for 24 h) were co-incubated with DiI-labeled exosomes isolated from A549 and H520 cells. (D) Fluorescence microscopy images of THP-1-M0 macrophages after co-incubation with PKH67-labeled exosomes (scale bar: 20 μm). (E-F) Western blot and quantification of EHF protein levels in THP-1-M0 macrophages after incubation with exosomes from A549 or H520 cells. Data are presented as mean ± SD. P < 0.05 denotes statistical significance.

Article Snippet: Human monocyte cell line (THP-1) (Catalog #: YC-D011) were purchased from Guangzhou Ubigene Biosciences Co., Ltd. (Guangzhou, China) and cultured in RPMI-1640 (Gibco) containing 10% FBS (Gibco).

Techniques: Derivative Assay, Isolation, Western Blot, Marker, Incubation, Labeling, Fluorescence, Microscopy

Journal: Regenerative Therapy

Article Title: Lung cancer cell-derived exosomal EHF drives M2 macrophage polarization via transcriptional activation of RNF41 to promote tumor progression

doi: 10.1016/j.reth.2026.101104

Figure Lengend Snippet: Exosomal EHF reprograms THP-1-M0 macrophages to promote lung cancer cell malignancy and angiogenesis via paracrine signaling. (A) Schematic of the paracrine co-culture system: Exosomes from control (Ctrl) or EHF-knockdown (KD-EHF) A549/H520 cells were co-incubated with THP-1-M0 macrophages. The medium from these macrophages was used to treat A549/H520 cells or HUVECs. (B-E) THP-1-M0 macrophages were treated with three conditions: blank control (no exosome), exosomes from control-transfected A549/H520 cells (Exo Ctrl ), or exosomes from EHF-knockdown A549/H520 cells (Exo KD−EHF ). (B–C) Flow cytometry analysis of CD206 + M2 macrophage proportions in THP-1-M0 cells. (D-E) ELISA measurement of M2-associated cytokines (TGF-β1, IL-10, and VEGFA). (F-M) The medium from exosome treated macrophages was then used to incubate A549/H520 cells or HUVECs. (F-G) EdU staining and quantification of EdU + proliferating A549 and H520 cells. (H–I) Flow cytometric analysis of apoptosis in A549 and H520 cells. (J-K) Transwell migration assays and quantification of migrated A549 and H520 cells. (L-M) Tube formation assays and quantification of tube numbers in HUVECs. Data are presented as mean ± SD. P < 0.05 denotes statistical significance.

Article Snippet: Human monocyte cell line (THP-1) (Catalog #: YC-D011) were purchased from Guangzhou Ubigene Biosciences Co., Ltd. (Guangzhou, China) and cultured in RPMI-1640 (Gibco) containing 10% FBS (Gibco).

Techniques: Co-Culture Assay, Control, Knockdown, Incubation, Transfection, Flow Cytometry, Enzyme-linked Immunosorbent Assay, Staining, Migration

Journal: Regenerative Therapy

Article Title: Lung cancer cell-derived exosomal EHF drives M2 macrophage polarization via transcriptional activation of RNF41 to promote tumor progression

doi: 10.1016/j.reth.2026.101104

Figure Lengend Snippet: EHF transcriptionally activates RNF41 and mediates its intercellular transfer via exosomes. (A) JASPAR database prediction of the EHF-binding motif within the RNF41 promoter region, localized to positions −828/-821. (B) ChIP-qPCR analysis in THP-1-M0 macrophages. (C) Dual-luciferase reporter assays in 293T cells. (D) WB analysis of EHF protein levels in THP-1-M0 macrophages with EHF knockdown (KD-EHF) or control (Ctrl). (E-F) Western blot and qRT-PCR analysis of RNF41 protein and mRNA levels in THP-1-M0 macrophages with KD-EHF or Ctrl. (G-H) THP-1-M0 macrophages were treated with culture medium (Blank), control exosomes (Ctrl Exo, from A549/H520 cells), or EHF-knockdown exosomes (KD-EHF Exo, from A549/H520-KD-EHF cells). RNF41 protein levels were detected by Western blot. Data are presented as mean ± SD. P < 0.05 denotes statistical significance.

Article Snippet: Human monocyte cell line (THP-1) (Catalog #: YC-D011) were purchased from Guangzhou Ubigene Biosciences Co., Ltd. (Guangzhou, China) and cultured in RPMI-1640 (Gibco) containing 10% FBS (Gibco).

Techniques: Binding Assay, ChIP-qPCR, Luciferase, Knockdown, Control, Western Blot, Quantitative RT-PCR

Journal: Regenerative Therapy

Article Title: Lung cancer cell-derived exosomal EHF drives M2 macrophage polarization via transcriptional activation of RNF41 to promote tumor progression

doi: 10.1016/j.reth.2026.101104

Figure Lengend Snippet: RNF41 mediates exosomal EHF-driven pro-tumorigenic effects in lung cancer. (A) Schematic of the rescue co-culture system: THP-1-M0 macrophages (control or RNF41-overexpressing) were incubated with exosomes from control (Exo Ctrl ) or EHF-knockdown (Exo KD−EHF ) A549/H520 cells. The medium from these co-cultures was used to treat A549/H520 cells or HUVECs. (B–C) Flow cytometry was used to analyze the proportion of CD206 + M2-polarized macrophages in THP-1-M0 cells. (D-E) ELISA was employed to quantify the levels of M2-associated cytokines (TGF-β1, IL-10, and VEGFA) in the CM from macrophages. (F-G) EdU assays were carried out to measure the proliferation of A549 and H520 cells. (H–I) Annexin V/PI flow cytometry was used to assess the apoptosis of A549 and H520 cells treated with CM. (J-K) Transwell assays were performed to evaluate the migration of A549 and H520 cells. (L-M) Tube formation assays were conducted on HUVECs treated with CM. Data are presented as mean ± SD. P < 0.05 denotes statistical significance.

Article Snippet: Human monocyte cell line (THP-1) (Catalog #: YC-D011) were purchased from Guangzhou Ubigene Biosciences Co., Ltd. (Guangzhou, China) and cultured in RPMI-1640 (Gibco) containing 10% FBS (Gibco).

Techniques: Co-Culture Assay, Control, Incubation, Knockdown, Flow Cytometry, Enzyme-linked Immunosorbent Assay, Migration

Journal: The Journal of Biological Chemistry

Article Title: Improvement in binding and function of a monoclonal antibody against Shigella flexneri 3a O-antigen via phage display and whole-cell in-solution panning

doi: 10.1016/j.jbc.2026.111405

Figure Lengend Snippet: Functional analysis of hFlex3a2_v2 variants against S. flexneri 3a. A , L-ABA assay comparing the ability of single hFlex3a2_v2 variants or ( B ) combinatorial hFlex3a2_v2 variants and a non-binding control to coordinate complement-mediated bacteriolysis of S. flexneri 3a. L-ABA is the average of three biological replicates each performed in duplicate. L-ABAs in panels ( A ) and ( B ) were performed in parallel but graphed separately for clarity. hFlex3a2_v2 and control mAb data is the same in these two panels. p values in were calculated on logIC 50 values by repeated measure Anova with Dunnett’s correction for multiple comparisons. C , OPA assay measures the percentage of THP-1 cells that have phagocytosed S. flexneri 3a that was opsonized with 30 nM of the indicated mAb. This experiment is the result of five biological replicates each performed in duplicate, and p values were calculated by repeated measure ANOVA with Dunnett’s correction for multiple comparisons. Error bars represent standard deviation. D , Invasion assay shows the percent invasion of S. flexneri 3a incubated with 1 μM of the indicated mAb normalized to the invasion rate of S. flexneri 3a in PBS. This experiment is the result of four biological replicates each performed in duplicate, and p -values were calculated by repeated measure ANOVA with Dunnett’s correction for multiple comparisons. Error bars represent standard deviation. For all data, p values can be found in and those < 0.5 are denoted by a ∗. ( E ) A cartoon depicting the top-ranked Alphafold model of hFlex3a2_v2 (pLDDT = 96. 2, pTM = 0.929, and ipTM = 0.909). The VL is shown in light gray, and the VH is shown in dark gray. CDRL loops are colored orange, and residues where variants were selectively enriched against S. flexneri 3a following panning against S. flexneri 3a and S. sonnei and were experimentally characterized are colored green , while H34 (not selectively enriched) is shown in pink .

Article Snippet: THP-1 cell lines were obtained from ATCC.

Techniques: Functional Assay, Binding Assay, Control, OPA Assay, Standard Deviation, Invasion Assay, Incubation

Journal: The Journal of Biological Chemistry

Article Title: Improvement in binding and function of a monoclonal antibody against Shigella flexneri 3a O-antigen via phage display and whole-cell in-solution panning

doi: 10.1016/j.jbc.2026.111405

Figure Lengend Snippet: Functional analysis of hFlex3a2_v2 variants against S. flexneri 3b. Binding curves from a titration ELISA against S. flexneri 3b OMVs (2.5 μg/ml) show relative binding potencies of hFlex3a2_v2 ( A ) single amino acid variants or ( B ) hFlex3a2_v2 combinatorial amino acid variants. ELISA binding curves are the average of three independent replicates each performed in duplicate. ELISAs in panels ( A ) and ( B ) were performed in parallel but graphed separately for clarity. hFlex3a2_v2 and control mAb data is the same in these two panels. ( C ) L-ABA assay comparing the ability of single hFlex3a2_v2 variants or ( D ) combinatorial hFlex3a2_v2 variants and a non-binding control to coordinate complement-mediated bacteriolysis of S. flexneri 3b. L-ABA is the average of three biological replicates each performed in duplicate. L-ABAs in panels ( C ) and ( D ) were performed in parallel but graphed separately for clarity. hFlex3a2_v2 and control mAb data is the same in these two panels. p -values were calculated on logIC 50 values by repeated measure Anova with Dunnett’s correction for multiple comparisons ( E ) OPA assay measures the percent of THP-1 cells that have phagocytosed S. flexneri 3b that was opsonized with 30 nM of the indicated mAb. This experiment is the result of five biological replicates each performed in duplicate, and p -values were calculated by repeated measure ANOVA with Dunnett’s correction for multiple comparisons. Error bars represent standard deviation. F , invasion assay shows the percent invasion of S. flexneri 3b incubated with 1 μM of the indicated mAb normalized to the invasion rate of S. flexneri 3b in PBS. This experiment is the result of four biological replicates each performed in duplicate. Error bars represent standard deviation. For all plots, p values can be found in and those < 0.5 are denoted by a ∗.

Article Snippet: THP-1 cell lines were obtained from ATCC.

Techniques: Functional Assay, Binding Assay, Titration, Enzyme-linked Immunosorbent Assay, Control, OPA Assay, Standard Deviation, Invasion Assay, Incubation

Journal: Translational Oncology

Article Title: Unmasking FCGR2B as a high-grade serous ovarian cancer specific marker of immune suppression and tumor progression through multi-omics mining

doi: 10.1016/j.tranon.2026.102748

Figure Lengend Snippet: FCGR2B expression defines an immunosuppressive TME in HGSOC. (A) ssGSEA-based immune infiltration analysis across 28 immune cell types. Δ = High - Low. (B) CIBERSORT-based macrophage-focused immune deconvolution reveals distinct immune landscapes between FCGR2B -High and FCGR2B -Low tumors. (C) Fluorescence images (scale bar, 50 μm) of FCGR2B (red), CD14 (green), CD68 (yellow), and the corresponding merged image. (D-E) Quantitative analysis of FCGR2B co-localization with CD14 (D) and CD68 (E). Left: box plots comparing co-localization levels between FCGR2B-High and FCGR2B-Low tumors (defined by the median FCGR2B H-score; Wilcoxon rank-sum test). Right: scatter plots showing correlations between FCGR2B and marker H-scores across all HGSOC samples (Spearman’s rho). (F) FCGR2B protein expression in HGSOC cell lines (SKOV3 and OVCAR3) and the cell line THP-1, as detected by WB assays. (G) FCGR2B protein expression in THP-1 cells transfected with Si-FCGR2B or control siRNA, as detected by WB assays. (H) Effect of FCGR2B knockdown on macrophage M2 polarization (CD206⁺/CD68⁺) under SKOV3 (HGSOC) co-culture conditions, assessed by flow cytometry. (I) Effect of FCGR2B knockdown on macrophage phagocytosis (CD68⁺/CFSE⁺) of SKOV3 cells under co-culture conditions, analyzed by flow cytometry. Statistical significance was determined using a two-tailed unpaired Student’s t -test; exact P values are indicated within the figure ( P = 0.0001, t = 14.35 for H; P = 0.0013, t = 7.976 for I; n =3, df = 4).

Article Snippet: The HGSOC cell lines SK-OV-3 (ATCC® HTB-77TM) and OVCAR-3 (ATCC® HTB-161TM), as well as the human monocytic cell line THP-1 (ATCC® TIB-202TM), were obtained from the American Type Culture Collection and cultured under standard conditions at 37 °C with 5% CO2.

Techniques: Expressing, Fluorescence, Marker, Transfection, Control, Knockdown, Co-Culture Assay, Flow Cytometry, Two Tailed Test

Journal: Journal of Immunology Research

Article Title: Aspergillus fumigatus Gliotoxin Inhibits LC3‐Associated Phagocytosis in Macrophages in a Calcium‐Dependent Manner

doi: 10.1155/jimr/5542735

Figure Lengend Snippet: GT attenuates LAP and in THP1 cells. (A and B) Western blot analysis of LC3‐II and SQSTM 1/p62 in THP1‐derived macrophages treated with gliotoxin (GT, 25 ng/mL) for the indicated time points (0–120 min). LC3‐II and SQSTM 1/p62 accumulation were reduced in a time‐dependent manner. (C and D) THP1‐derived macrophages were exposed to increasing concentrations of GT (0–100 ng/mL) for 2 h. LC3‐II and SQSTM 1/p62 levels decreased in a dose‐dependent fashion. Densitometric quantification of LC3‐II was normalized to GAPDH. (E) Representative confocal micrographs showing LC3 fluorescence (red) in THP1 cells treated with 25 ng/mL GT for 2 h. Nuclei were counterstained with DAPI (blue). GT markedly reduced LC3 puncta formation compared with untreated controls. All experiments were independently repeated at least three times. GAPDH loading controls in (A) and (C) were derived from the same membrane.

Article Snippet: Cell lines: The THP‐1 cell line (human acute monocytic leukemia cells) and the mh‐s cell line (mouse alveolar macrophages) were obtained from ATCC (America Type Culture Collection, TIB‐202, CRL‐2019) and cultured in RPMI‐1640 (GIBCO, Germany) supplemented with 10% heat‐inactivated fetal calf serum, 100 U/mL streptomycin, and 100 U/mL penicillin at 37°C in an atmosphere of 5% CO 2.

Techniques: Western Blot, Derivative Assay, Fluorescence, Membrane

Journal: Journal of Immunology Research

Article Title: Aspergillus fumigatus Gliotoxin Inhibits LC3‐Associated Phagocytosis in Macrophages in a Calcium‐Dependent Manner

doi: 10.1155/jimr/5542735

Figure Lengend Snippet: GT attenuates A. fumigatus ‐induced LAP in THP1 cells. (A and C) THP1‐derived macrophages were infected with WT, gliP Δ, or gliP R swollen conidia of A. fumigatus (MOI = 5) for 2 h. Western blot and quantification of LC3‐I/II showed that LAP activation was significantly attenuated in cells infected with WT or gliP R A. fumigatus compared to gliP Δ strains. (B and D) THP1‐derived macrophages were infected with WT and gliP Δ conidia (MOI = 5) in the presence or absence of GT (25 ng/mL) for 2 h. GT was added simultaneously with conidia. Western blot and confocal microscopy revealed that GT suppressed the LC3‐II conversion and SQSTM1/p62 expression induced by gliP Δ infection. All experiments were performed independently at least three times. GAPDH loading controls in (A) and (C) were derived from the same membrane.

Article Snippet: Cell lines: The THP‐1 cell line (human acute monocytic leukemia cells) and the mh‐s cell line (mouse alveolar macrophages) were obtained from ATCC (America Type Culture Collection, TIB‐202, CRL‐2019) and cultured in RPMI‐1640 (GIBCO, Germany) supplemented with 10% heat‐inactivated fetal calf serum, 100 U/mL streptomycin, and 100 U/mL penicillin at 37°C in an atmosphere of 5% CO 2.

Techniques: Derivative Assay, Infection, Western Blot, Activation Assay, Confocal Microscopy, Expressing, Membrane

Journal: Journal of Immunology Research

Article Title: Aspergillus fumigatus Gliotoxin Inhibits LC3‐Associated Phagocytosis in Macrophages in a Calcium‐Dependent Manner

doi: 10.1155/jimr/5542735

Figure Lengend Snippet: GT inhibits LAP in THP1 cells via a MAPK10‐independent manner. (A) Heatmap of differentially expressed genes (DEGs) in THP1 cells treated with gliP Δ compared to WT (MOI = 5, 2 h). The color scale represents log 2 fold changes in gene expression relative to control cells. (B) MAPK10 protein levels in THP1 cells infected with wWT, gliP Δ or gliP R swollen conidia of A. fumigatus (MOI = 5) for 2 h. (C) Western blot analysis of MAPK10 expression in THP1 macrophages after GT (25 ng/mL, 2 h) treatment. (D and E) THP1 cells were transfected with MAPK10 siRNA for 48 h or pretreated with JNK inhibitor (SP600125, 20 μM, 1 h), followed by GT (25 ng/mL) exposure for 2 h. Western blot analysis and densitometric quantification of LC3, SQSTM1/p62, and Rubicon showed that GT inhibited LAP regardless of MAPK10 or JNK inhibition. All experiments were performed independently at least three times.

Article Snippet: Cell lines: The THP‐1 cell line (human acute monocytic leukemia cells) and the mh‐s cell line (mouse alveolar macrophages) were obtained from ATCC (America Type Culture Collection, TIB‐202, CRL‐2019) and cultured in RPMI‐1640 (GIBCO, Germany) supplemented with 10% heat‐inactivated fetal calf serum, 100 U/mL streptomycin, and 100 U/mL penicillin at 37°C in an atmosphere of 5% CO 2.

Techniques: Gene Expression, Control, Infection, Western Blot, Expressing, Transfection, Inhibition

Journal: Journal of Immunology Research

Article Title: Aspergillus fumigatus Gliotoxin Inhibits LC3‐Associated Phagocytosis in Macrophages in a Calcium‐Dependent Manner

doi: 10.1155/jimr/5542735

Figure Lengend Snippet: Calcium involved in the decreased LAP of THP1 cells exposed to GT. (A and B) THP1 macrophages were treated with GT (25 ng/mL) for 2 h. Intracellular calcium concentrations were determined using a fluorescence microplate reader and flow cytometry (excitation 494 nm, emission 516 nm). (C) THP1 cells were treated with GT (25 ng/mL, 2 h), calcium chelator W7 (10 μM, 1 h), or both. Intracellular ROS levels were measured fluorometrically to assess the contribution of calcium signaling to GT‐induced oxidative stress. (D) THP1 macrophages were treated with GT (25 ng/mL, 2 h), W7 (10 μM, 1 h), or both. Western blot analysis and densitometric quantification of LC3‐II, SQSTM1/p62, and Rubicon expression demonstrated that calcium inhibition partially reversed GT‐induced suppression of LAP. All experiments were independently repeated at least three times.

Article Snippet: Cell lines: The THP‐1 cell line (human acute monocytic leukemia cells) and the mh‐s cell line (mouse alveolar macrophages) were obtained from ATCC (America Type Culture Collection, TIB‐202, CRL‐2019) and cultured in RPMI‐1640 (GIBCO, Germany) supplemented with 10% heat‐inactivated fetal calf serum, 100 U/mL streptomycin, and 100 U/mL penicillin at 37°C in an atmosphere of 5% CO 2.

Techniques: Fluorescence, Flow Cytometry, Western Blot, Expressing, Inhibition

Journal: Scientific Reports

Article Title: Renal denervation attenuates cardiac fibrosis and improves left ventricular function in rats with myocardial infarction

doi: 10.1038/s41598-026-50195-w

Figure Lengend Snippet: β-adrenergic stimulation of macrophages causes upregulation of profibrotic and downregulation of inflammatory genes via β1-adrenoceptor. ( a ) Experimental setup: THP-1 cells were differentiated to macrophages by stimulation with 50 nM PMA for 24 h, followed by stimulation with β-adrenoceptor agonist ISO or PBS (CTRL) in the presence or absence of β1-adrenoceptor antagonist CGP or β2-adrenoceptor antagonist ICI over a period of 72 h with daily stimulations. Cells were harvested and gene expression analysis was conducted for macrophage phenotype markers. ( b ) Arginase 1 (ARG1) and CTGF genes were higher in ISO compared to CTRL. Compared to ISO, expression was smaller in CGP + ISO but not ICI + ISO. ( c ) CXC motif chemokine ligand 10 (CXCL10) and CD80 gene expression were significantly smaller in ISO compared to CTRL. Compared to ISO, expression was higher in CGP + ISO but not ICI + ISO. Data are obtained as duplicates from at least three independent experiments and shown as mean ± SEM. P-value was determined using ANOVA with Tukey’s test for multiple comparisons. CTRL=Control group (Stimulation with phosphate-buffered saline (PBS)); ISO=Isoprenaline stimulation group; CGP = β1-adrenoceptor inhibitor; ICI = β2-adrenoceptor inhibitor; mRNA=messenger ribonucleic acid; ARG1 = Arginase 1; CTGF=Connective tissue growth factor; CXCL10 = C-X-C motif chemokine 10; CD=cluster of differentiation.

Article Snippet: The human monocytic cell line THP-1 was purchased from ATCC (Wesel, Germany).

Techniques: Gene Expression, Expressing, Control, Saline