Journal: Journal of Oncology

Article Title: Cancer-Associated Fibroblast-Derived Exosomal miRNA-320a Promotes Macrophage M2 Polarization In Vitro by Regulating PTEN/PI3K γ Signaling in Pancreatic Cancer

doi: 10.1155/2022/9514697

Figure Lengend Snippet: CAFs-derived conditioned medium (CAFs-CM) facilitated macrophage M2 polarization. (a) qPCR analysis of mRNA expression of classic M2 signature markers (CD163, CD206, and IL-10) and classic M1 signature markers (HLA-DR and iNOS) in U937 cells cultured with HPDE6-C7-CM, PANC-CM, NF3-CM, NF6-CM, CAF3-CM, or CAF6-CM. (b) qPCR analysis of mRNA expression of CD163, CD206, and IL-10 in U937 cells cultured with NFs-CM, CAFs-CM, or CAFs/GW4869-CM. (c) Western blot analysis of CD163, CD206, and IL-10 protein expression in U937 cells cultured with NFs-CM, CAFs-CM, or CAFs/GW4869-CM. (d) Quantitative analysis of CD163, CD206, and IL-10 protein expression in U937 cells cultured with NFs-CM, CAFs-CM, or CAFs/GW4869-CM. (e) The effect of NFs-CM, CAFs-CM, or CAFs/GW4869-CM on proliferation of pancreatic cancer cells by CCK-8 assay. (f) The effect of NFs-CM, CAFs-CM, or CAFs/GW4869-CM on pancreatic cancer cells invasion was determined by the Transwell assay. Representative photographs (magnification, 100) and the number of invaded cells are displayed. ∗∗ p < 0.01. # vs CAFs-CM. qPCR, quantitative real-time PCR; CM, conditioned medium; NFs, normal fibroblasts; CAFs, cancer-associated fibroblasts.

Article Snippet: Human monocytic cell line U937 was purchased from Merck Millipore (Temecula, California, USA) and cultured in DMEM supplemented with 10% FBS, 1% penicillin/streptomycin, and 10 nM 12-o-tetradecanoyl-phorbol-13-acetate (TPA).

Techniques: Derivative Assay, Expressing, Cell Culture, Western Blot, CCK-8 Assay, Transwell Assay, Real-time Polymerase Chain Reaction

Journal: Journal of Oncology

Article Title: Cancer-Associated Fibroblast-Derived Exosomal miRNA-320a Promotes Macrophage M2 Polarization In Vitro by Regulating PTEN/PI3K γ Signaling in Pancreatic Cancer

doi: 10.1155/2022/9514697

Figure Lengend Snippet: CAFs-derived exosomes (CAFs-Exo) facilitated macrophage M2 polarization. (a) Electron micrograph analysis of exosomes collected from CAFs-CM (bar, 500 nm). (b) Size distribution and concentration range characterization of exosomes collected from CAFs-CM assayed with qNano. (c) Western blot analysis of protein expression of CD63, CD81, and HSP70 (markers of exosomes) in exosomes collected from CAFs-CM assayed. (d) qPCR analysis of CD163, CD206, and IL-10 mRNA expression in U937 cells cultured with NFs-Exo or CAFs-Exo. (e) Western blot analysis of protein expression of CD163, CD206, and IL-10 in U937 cells cultured with NFs-Exo or CAFs-Exo. (f) Quantitative analysis of CD163, CD206, and IL-10 protein expression in U937 cells cultured with NFs-Exo or CAFs-Exo. ∗∗ p < 0.01. qPCR, quantitative real-time PCR; CM, conditioned medium; NFs, normal fibroblasts; CAFs, cancer-associated fibroblasts; Exo, exosome.

Article Snippet: Human monocytic cell line U937 was purchased from Merck Millipore (Temecula, California, USA) and cultured in DMEM supplemented with 10% FBS, 1% penicillin/streptomycin, and 10 nM 12-o-tetradecanoyl-phorbol-13-acetate (TPA).

Techniques: Derivative Assay, Concentration Assay, Western Blot, Expressing, Cell Culture, Real-time Polymerase Chain Reaction

Journal: Journal of Oncology

Article Title: Cancer-Associated Fibroblast-Derived Exosomal miRNA-320a Promotes Macrophage M2 Polarization In Vitro by Regulating PTEN/PI3K γ Signaling in Pancreatic Cancer

doi: 10.1155/2022/9514697

Figure Lengend Snippet: Exosomes mediated the transfer of miRNA-320a from CAFs to macrophages. qPCR analysis of miR-106b, miR-148a, miR-125b, miR-320c, miR-320a, miR-1285, miR-422a, miR-29a, and miR-378d mRNA expression in NFs and CAFs (a) or NFs-Exo and CAFs-Exo (b). (c, d) qPCR analysis of miR-320a mRNA expression in 7 pancreatic cancer tissues-derived CAFs and 7 pancreatic cancer tissues-derived NFs or 7 pancreatic cancer tissues-derived CAFs-Exo and 7 pancreatic cancer tissues-derived NFs-Exo. (e–g) qPCR analysis of miR-320a mRNA expression in miRNA-320a-overexpressed CAFs (e), corresponding CAFs-Exo (f), and U937 cells after treatment with miRNA-320a-overexpressed CAFs-Exo (g). (h) Fluorescence microscope analysis of FAM-tagged miRNA-320a in U937cells treated with CAFs-Exo/FAM-miRNA-320a. ∗∗ p < 0.01. qPCR, quantitative real-time PCR; CM, conditioned medium; NFs, normal fibroblasts; CAFs, cancer-associated fibroblasts; Exo, exosome; CAFs-Exo/FAM-miRNA-320a, FAM-miRNA-320a-overexpressed CAFs-Exo.

Article Snippet: Human monocytic cell line U937 was purchased from Merck Millipore (Temecula, California, USA) and cultured in DMEM supplemented with 10% FBS, 1% penicillin/streptomycin, and 10 nM 12-o-tetradecanoyl-phorbol-13-acetate (TPA).

Techniques: Expressing, Derivative Assay, Fluorescence, Microscopy, Real-time Polymerase Chain Reaction

Journal: Journal of Oncology

Article Title: Cancer-Associated Fibroblast-Derived Exosomal miRNA-320a Promotes Macrophage M2 Polarization In Vitro by Regulating PTEN/PI3K γ Signaling in Pancreatic Cancer

doi: 10.1155/2022/9514697

Figure Lengend Snippet: miRNA-320a facilitated macrophage M2 polarization. qPCR analysis of classic M2 signature markers (CD163, CD206, and IL-10) and classic M1 signature markers (HLA-DR and iNOS) mRNA expression in U937 cells cultured with miRNA-320a mimics (a) or inhibitor (b). Western blot (c) and quantitative analysis (d) of CD163, CD206, and IL-10 protein expression in U937 cells cultured with miRNA-320a mimics. (e) The effect of U937/miR-320a-CM on pancreatic cancer cell proliferation by CCK-8 assay. (f, g) The effect of U937/miR-320a-CM on pancreatic cancer cells invasion was determined by the Transwell assay. Representative photographs (magnification, 100) and the number of invaded cells are displayed. ∗∗ p < 0.01. qPCR, quantitative real-time PCR; CM, conditioned medium; NFs, normal fibroblasts; CAFs, cancer-associated fibroblasts; U937/miR-320a-CM, miRNA-320a-overexpressed U937-derived CM.

Article Snippet: Human monocytic cell line U937 was purchased from Merck Millipore (Temecula, California, USA) and cultured in DMEM supplemented with 10% FBS, 1% penicillin/streptomycin, and 10 nM 12-o-tetradecanoyl-phorbol-13-acetate (TPA).

Techniques: Expressing, Cell Culture, Western Blot, CCK-8 Assay, Transwell Assay, Real-time Polymerase Chain Reaction, Derivative Assay

Journal: Journal of Oncology

Article Title: Cancer-Associated Fibroblast-Derived Exosomal miRNA-320a Promotes Macrophage M2 Polarization In Vitro by Regulating PTEN/PI3K γ Signaling in Pancreatic Cancer

doi: 10.1155/2022/9514697

Figure Lengend Snippet: miRNA-320a functions by targeting PTEN/PI3K γ signaling. (a) Schematic representation of the miR-320a site in PTEN-3′UTR. (b, c) Luciferase activity was assayed in PCa cells co-transfected with miR-320a and luciferase reporters containing PTEN-3′UTR. Data are presented as the relative ratio of firefly luciferase activity to renilla luciferase activity. Western blot (d) and quantitative analysis (e) of PTEN, PI3K γ , AKT, and p-AKT protein expression in miRNA-320a-overexpressed U937 cells. Western blot (f) and quantitative analysis (g) of CD163 and CD206 protein expression in U937 cells overexpressed with miRNA-320a in the presence or absence of PI3K γ siRNA. ∗∗ p < 0.01. # vs miR-320a mimic group. qPCR, quantitative real-time PCR.

Article Snippet: Human monocytic cell line U937 was purchased from Merck Millipore (Temecula, California, USA) and cultured in DMEM supplemented with 10% FBS, 1% penicillin/streptomycin, and 10 nM 12-o-tetradecanoyl-phorbol-13-acetate (TPA).

Techniques: Luciferase, Activity Assay, Transfection, Western Blot, Expressing, Real-time Polymerase Chain Reaction

Journal: Evidence-based Complementary and Alternative Medicine : eCAM

Article Title: Synergistic Antimycobacterial Actions of Knowltonia vesicatoria (L.f) Sims

doi: 10.1155/2012/808979

Figure Lengend Snippet: Undifferentiated (A) and differentiated (B) U937 cells as viewed under a light microscope (100x magnification).

Article Snippet: The histiocytic lymphoma cell line, U937, was obtained from Highveld Biological (Pty) (Ltd.) (Sandringham, South Africa) and maintained in complete RPMI 1640 medium (pH 7.2) (Sigma-Aldrich Chemical Co., South Africa), supplemented with 10% heat-inactivated foetal calf serum (FCS), 2 mM L-glutamine, and a 0.1% antimicrobial solution (pennicilin, streptomycin, and an antifungal. fungizone).

Techniques: Light Microscopy

Journal: Evidence-based Complementary and Alternative Medicine : eCAM

Article Title: Synergistic Antimycobacterial Actions of Knowltonia vesicatoria (L.f) Sims

doi: 10.1155/2012/808979

Figure Lengend Snippet: Antimycobacterial activity against M. tuberculosis and cytotoxicity on U937 cells of test samples.

Article Snippet: The histiocytic lymphoma cell line, U937, was obtained from Highveld Biological (Pty) (Ltd.) (Sandringham, South Africa) and maintained in complete RPMI 1640 medium (pH 7.2) (Sigma-Aldrich Chemical Co., South Africa), supplemented with 10% heat-inactivated foetal calf serum (FCS), 2 mM L-glutamine, and a 0.1% antimicrobial solution (pennicilin, streptomycin, and an antifungal. fungizone).

Techniques: Activity Assay

Journal: BMB Reports

Article Title: Diesel exhaust particles disrupt blood–retina barrier integrity via TLR2 and TLR4 activation

doi: 10.5483/BMBRep.2025-0013

Figure Lengend Snippet: TNF-α and IL-1β mRNA expression in U937 human macrophages at 6, 24, and 48 h after DEP exposure followed by washing. (A) Schematic representation of the experimental procedure. (B, C) TNF-α and IL-1β mRNA expression. TNF-α (B) and IL-1β (C) mRNA levels were significantly elevated at each time point following DEP exposure compared with those in the control. *P < 0.05 vs. control.

Article Snippet: U937 human macrophages (Korean Cell Line Bank, Seoul, Korea) were cultured in Dulbecco’s Modified Eagle Medium (DMEM) supplemented with 10% heat-inactivated fetal bovine serum (FBS, Gibco, MA, USA) for 3 days at 37°C in a 5% CO 2 incubator.

Techniques: Expressing, Control

Journal: BMB Reports

Article Title: Diesel exhaust particles disrupt blood–retina barrier integrity via TLR2 and TLR4 activation

doi: 10.5483/BMBRep.2025-0013

Figure Lengend Snippet: Claudin-5 and ZO-1 expression and permeability in human retinal endothelial cells (HRECs) treated with conditioned media from U937 macrophages exposed to DEP. (A) Schematic representation of the experimental procedure. (B) Immunocytochemical staining. Claudin-5 and ZO-1 expression in HRECs was markedly reduced after treatment with conditioned media from DEP-exposed U937 macrophages. (C, D) Western blot analysis. Claudin-5 (C) and ZO-1 (D) protein levels in HRECs were significantly decreased following treatment with DEP-exposed U937-conditioned media compared with the control. (E) TEER permeability assay. Permeability in HRECs was significantly increased after treatment with DEP-exposed U937-conditioned media compared with the control. Scale bar = 30 μm, *P < 0.05 vs. control.

Article Snippet: U937 human macrophages (Korean Cell Line Bank, Seoul, Korea) were cultured in Dulbecco’s Modified Eagle Medium (DMEM) supplemented with 10% heat-inactivated fetal bovine serum (FBS, Gibco, MA, USA) for 3 days at 37°C in a 5% CO 2 incubator.

Techniques: Expressing, Permeability, Staining, Western Blot, Control

Journal: BMB Reports

Article Title: Diesel exhaust particles disrupt blood–retina barrier integrity via TLR2 and TLR4 activation

doi: 10.5483/BMBRep.2025-0013

Figure Lengend Snippet: Expression levels of TLR2/TLR4 and TNF-α/IL-1β mRNA in U937 macrophages exposed to DEPs with TLR2 (C29) or TLR4 (TAK242) inhibitors. (A) Schematic representation of the experimental procedure. (B, C) TLR2 and TLR4 expression levels. Compared with those in the control, TLR2 (B) and TLR4 (C) levels in U937 macrophages were significantly elevated following DEP exposure. TLR2 and TLR4 expression levels were significantly restored following treatment with C29 and TAK242 before DEP exposure, respectively. (D, E) TNF-α and IL-1β mRNA expression. TNF-α (D) and IL-1β (E) mRNA levels were significantly lower in U937 macrophages treated with C29 or TAK242 before DEP exposure than in cells that received DEP exposure alone without TLR inhibitors. *P < 0.05 vs. control, † P < 0.05 vs. DEP.

Article Snippet: U937 human macrophages (Korean Cell Line Bank, Seoul, Korea) were cultured in Dulbecco’s Modified Eagle Medium (DMEM) supplemented with 10% heat-inactivated fetal bovine serum (FBS, Gibco, MA, USA) for 3 days at 37°C in a 5% CO 2 incubator.

Techniques: Expressing, Control

Journal: BMB Reports

Article Title: Diesel exhaust particles disrupt blood–retina barrier integrity via TLR2 and TLR4 activation

doi: 10.5483/BMBRep.2025-0013

Figure Lengend Snippet: Claudin-5 and ZO-1 expression and permeability in HRECs treated with conditioned media from U937 macrophages treated with C29 or TAK242 before exposure to DEPs. (A) Schematic representation of the experimental procedure. (B) Immunocytochemical staining. Claudin-5 and ZO-1 expression was markedly preserved in HRECs cultured in conditioned media from U937 macrophages treated with C29 or TAK242 before DEP exposure, than in HRECs that received DEP exposure alone without pretreatment with TLR inhibitors. (C, D) Western blot analysis. Claudin-5 (C) and ZO-1 (D) protein levels in HRECs were significantly restored following treatment with conditioned media from U937 macrophages treated with C29 or TAK242 before DEP exposure. (E) TEER permeability assay. Permeability was significantly lower in HRECs treated with conditioned media from U937 macrophages treated with C29 or TAK242 before DEP exposure, than in HRECs that received DEP exposure alone without pretreatment with TLR inhibitors. Scale bar = 30 μm, *P < 0.05 vs. control, † P < 0.05 vs. DEP.

Article Snippet: U937 human macrophages (Korean Cell Line Bank, Seoul, Korea) were cultured in Dulbecco’s Modified Eagle Medium (DMEM) supplemented with 10% heat-inactivated fetal bovine serum (FBS, Gibco, MA, USA) for 3 days at 37°C in a 5% CO 2 incubator.

Techniques: Expressing, Permeability, Staining, Cell Culture, Western Blot, Control