Journal: PLOS ONE

Article Title: Monocyte-cancer cell fusion is mediated by phosphatidylserine—CD36 receptor interaction and induced by ionizing radiation

doi: 10.1371/journal.pone.0311027

Figure Lengend Snippet: Fluorescent microscopy of CellTrace and CD36 staining A) MCF-7 cells stained with CellTrace Far red B) THP-1 cells stained with CellTrace Violet C) THP-1 cells stained with CD36 antibodies. All images were taken with x40 magnification.

Article Snippet: The cell cultures were washed in PBS, incubated with Human TruStain FcX TM (Fc Receptor Blocking Solution) (BioLegend, 422302, San Diego, USA) for 10 minutes at 4°C, 1:20 ratio to Cell Staining Buffer (Nordic Biosite, ASB-022501L, Täby, Sweden), followed by staining with 5 μg/ml Anti-Human CD36/SCARB3 Antibody (Phycoerythrin (PE)-conjugated) (Rabbit monoclonal IgG antibody, Sino Biological, 10752-R001-P, Beijing, China), or 50 μg/ml Rabbit IgG Isotype Control, PE-conjugated (Bioss, bs-0295P-PE, Massachusetts, USA) for 30 minutes in darkness at 4°C.

Techniques: Microscopy, Staining

Journal: PLOS ONE

Article Title: Monocyte-cancer cell fusion is mediated by phosphatidylserine—CD36 receptor interaction and induced by ionizing radiation

doi: 10.1371/journal.pone.0311027

Figure Lengend Snippet: Panel A shows THP-1 and MCF-7 cells cultured in separate flasks and treated with 0, 2.5, 5, and 10 Gy radiation. After that, the cells were analyzed by flow cytometry to detect the proportion of cells expressing PS and CD36. Panel B shows a cell fusion experiment where THP-1 cells (labelled with CellTrace Violet) and MCF-7 cells (labelled with CellTracer Far Red) are grown in the same flask and treated with 0 and 5 Gy gamma radiation. After 2 days of co-culture, the cells are sorted with FACS Aria III. Cells co-expressing CellTrace Violet and Far Red are assessed as THP1-/MCF-5 tumor hybrid cells.

Article Snippet: The cell cultures were washed in PBS, incubated with Human TruStain FcX TM (Fc Receptor Blocking Solution) (BioLegend, 422302, San Diego, USA) for 10 minutes at 4°C, 1:20 ratio to Cell Staining Buffer (Nordic Biosite, ASB-022501L, Täby, Sweden), followed by staining with 5 μg/ml Anti-Human CD36/SCARB3 Antibody (Phycoerythrin (PE)-conjugated) (Rabbit monoclonal IgG antibody, Sino Biological, 10752-R001-P, Beijing, China), or 50 μg/ml Rabbit IgG Isotype Control, PE-conjugated (Bioss, bs-0295P-PE, Massachusetts, USA) for 30 minutes in darkness at 4°C.

Techniques: Cell Culture, Flow Cytometry, Expressing, Co-Culture Assay

Journal: PLOS ONE

Article Title: Monocyte-cancer cell fusion is mediated by phosphatidylserine—CD36 receptor interaction and induced by ionizing radiation

doi: 10.1371/journal.pone.0311027

Figure Lengend Snippet: The proportion of MCF-7 expressing PS increases in dose-dependent relation (C). No correlation of PS and CD36 expression in THP-1 cells nor CD36 expression in MCF-7 cells was found in relation to the dose of ionizing radiation.

Article Snippet: The cell cultures were washed in PBS, incubated with Human TruStain FcX TM (Fc Receptor Blocking Solution) (BioLegend, 422302, San Diego, USA) for 10 minutes at 4°C, 1:20 ratio to Cell Staining Buffer (Nordic Biosite, ASB-022501L, Täby, Sweden), followed by staining with 5 μg/ml Anti-Human CD36/SCARB3 Antibody (Phycoerythrin (PE)-conjugated) (Rabbit monoclonal IgG antibody, Sino Biological, 10752-R001-P, Beijing, China), or 50 μg/ml Rabbit IgG Isotype Control, PE-conjugated (Bioss, bs-0295P-PE, Massachusetts, USA) for 30 minutes in darkness at 4°C.

Techniques: Expressing

Journal: PLOS ONE

Article Title: Monocyte-cancer cell fusion is mediated by phosphatidylserine—CD36 receptor interaction and induced by ionizing radiation

doi: 10.1371/journal.pone.0311027

Figure Lengend Snippet: The proportion of THP-1/MCF-7 tumor hybrid cells in co-cultures supplemented with 40 μg/ml and 80 μg/ml CD36 antibody was 1.8% and 1.9%, respectively. In non-treated THP-1/MCF-7 co-cultures, the proportion of tumor hybrid cells (3.6%) was significantly higher compared to co-cultures treated with anti-CD36 antibody (P<0.001).

Article Snippet: The cell cultures were washed in PBS, incubated with Human TruStain FcX TM (Fc Receptor Blocking Solution) (BioLegend, 422302, San Diego, USA) for 10 minutes at 4°C, 1:20 ratio to Cell Staining Buffer (Nordic Biosite, ASB-022501L, Täby, Sweden), followed by staining with 5 μg/ml Anti-Human CD36/SCARB3 Antibody (Phycoerythrin (PE)-conjugated) (Rabbit monoclonal IgG antibody, Sino Biological, 10752-R001-P, Beijing, China), or 50 μg/ml Rabbit IgG Isotype Control, PE-conjugated (Bioss, bs-0295P-PE, Massachusetts, USA) for 30 minutes in darkness at 4°C.

Techniques:

Journal: Archives of Toxicology

Article Title: Role of miR-24-3p and miR-146a-5p in dendritic cells’ maturation process induced by contact sensitizers

doi: 10.1007/s00204-023-03542-z

Figure Lengend Snippet: Cell surface markers expression after iDCs pre-treatment with miR-24-3p inhibitor and mimic-miR-146a-5p. iDCs were exposed for 24 h to BB (2 μg/mL) and HCA (20 μg/mL) in presence or absence of the miR-24-3p inhibitor (50 nM) or miRNA mimic (5 nM). After 24, the expression of CD36, CD86, CD206, CCR7 and HLA-DR were evaluated. Results are expressed as stimulation index (treatment/control). Each value represents the mean of n = 3 independent experiments. Statistical analysis was performed with a Dunnett’s comparison test, with * ,# p < 0.05, ** ,## p < 0.01 and *** , ### p < 0.001 vs control cells (* was used for statistically significant increase expression; # was used for statistically significant decrease expression). Dotted line is set up at 1.0 (control)

Article Snippet: Supernatants were collected for cytokines assessment, and cells stained with specific FITC/PE/Per-CP-conjugated antibodies against CD36, CD40, CD80, CD86, CD206, CCR7 and HLA-DR (BD, Becton Dickinson and ImmunoTools) or with isotype control antibodies at 4 °C following supplier's instructions.

Techniques: Expressing, Control, Comparison

Journal: Archives of Toxicology

Article Title: Role of miR-24-3p and miR-146a-5p in dendritic cells’ maturation process induced by contact sensitizers

doi: 10.1007/s00204-023-03542-z

Figure Lengend Snippet: Cell surface markers expression after iDCs pre-treatment with miR-24-3p inhibitor and mimic-miR-146a-5p. iDCs were exposed for 24 h to BB (2 μg/mL) and HCA (20 μg/mL) in presence or absence of the miR-24-3p inhibitor (50 nM) or miRNA mimic (5 nM). After 24, the expression of CD36, CD86, CD206, CCR7 and HLA-DR were evaluated. Results are expressed as stimulation index (treatment/control). Each value represents the mean of n = 3 independent experiments. Statistical analysis was performed with a Dunnett’s comparison test, with * ,# p < 0.05, ** ,## p < 0.01 and *** , ### p < 0.001 vs control cells (* was used for statistically significant increase expression; # was used for statistically significant decrease expression). Dotted line is set up at 1.0 (control)

Article Snippet: Supernatants were collected for cytokines assessment, and cells stained with specific FITC/PE/Per-CP-conjugated antibodies against CD36, CD40, CD80, CD86, CD206, CCR7 and HLA-DR (BD, Becton Dickinson and ImmunoTools) or with isotype control antibodies at 4 °C following supplier's instructions.

Techniques: Expressing, Control, Comparison

Journal: Molecular Medicine

Article Title: Oxidized Low-Density Lipoprotein-Dependent Platelet-Derived Microvesicles Trigger Procoagulant Effects and Amplify Oxidative Stress

doi: 10.2119/molmed.2011.00295

Figure Lengend Snippet: CD36-dependent platelet activation in response to PMVs. (A–C) The MFI and percentage of PAC-1, as well as platelet aggregation, were enhanced significantly after incubation of CD36-positive platelets with PMVs (30 μg/mL) for 30 min at 22°C. Data are means ± SE (n = 3). *P < 0.05 compared with control. (D–F) PAC-1 expression or platelet aggregation with CD36-deficient platelet incubation in response to PMVs. Data are means ± SE (n = 4). *P < 0.05 compared with control.

Article Snippet: Phycoerythrin (PE)-conjugated anti-CD36 (clone CB38), PE-conjugated CD36 isotype control (clone G155-228), PE-cy™5-conjugated mouse anti-CD41a antibody (clone HIP8) and fluorescein isothiocyanate (FITC)-conjugated PAC-1 were from BD Biosciences/Pharmingen (San Jose, CA, USA).

Techniques: Activation Assay, Incubation, Expressing

Journal: Molecular Medicine

Article Title: Oxidized Low-Density Lipoprotein-Dependent Platelet-Derived Microvesicles Trigger Procoagulant Effects and Amplify Oxidative Stress

doi: 10.2119/molmed.2011.00295

Figure Lengend Snippet: Identification of CD36-positive and CD36-deficient platelets by flow cytometry. MFI of binding of CD36-positive and CD36-deficient platelets with PE-conjugated anti-CD36 antibody or isotype controls.

Article Snippet: Phycoerythrin (PE)-conjugated anti-CD36 (clone CB38), PE-conjugated CD36 isotype control (clone G155-228), PE-cy™5-conjugated mouse anti-CD41a antibody (clone HIP8) and fluorescein isothiocyanate (FITC)-conjugated PAC-1 were from BD Biosciences/Pharmingen (San Jose, CA, USA).

Techniques: Flow Cytometry, Binding Assay

Journal: Molecular Medicine

Article Title: Oxidized Low-Density Lipoprotein-Dependent Platelet-Derived Microvesicles Trigger Procoagulant Effects and Amplify Oxidative Stress

doi: 10.2119/molmed.2011.00295

Figure Lengend Snippet: CD36-, PS- and JNK-dependent platelet activation by PMVs. Platelets were incubated with 2 μg/mL FA6 (A,D), 2 μg/mL mouse IgG or 20 μmol/L SP600125 (C,E) before incubation with PMVs (30 μg/mL) and analyzed by flow cytometry with FITC-labeled PAC-1. In some studies, PMVs were first incubated with 20 μg/mL annexin V to mask PS (B,D). (A–C) Histograms represent MFI and graphs; (D–E) show percentage of PAC-1–positive platelets. Data are means ± SE from at least three separate experiments. *P < 0.05 compared with control; #P < 0.05 compared with PMV treatment.

Article Snippet: Phycoerythrin (PE)-conjugated anti-CD36 (clone CB38), PE-conjugated CD36 isotype control (clone G155-228), PE-cy™5-conjugated mouse anti-CD41a antibody (clone HIP8) and fluorescein isothiocyanate (FITC)-conjugated PAC-1 were from BD Biosciences/Pharmingen (San Jose, CA, USA).

Techniques: Activation Assay, Incubation, Flow Cytometry, Labeling

Journal: Molecular Medicine

Article Title: Oxidized Low-Density Lipoprotein-Dependent Platelet-Derived Microvesicles Trigger Procoagulant Effects and Amplify Oxidative Stress

doi: 10.2119/molmed.2011.00295

Figure Lengend Snippet: (A–B) Expression of CD36 detected by flow cytometry. Preincubation of platelets with FA6, IgG, annexin V or SP600125 to examine binding of PE-conjugated CD36 antibody to platelets. (C–D) P-selection secretion determined by ELISA. The secretion of P-selection with blocking of CD36 or PS (C); the effect of JNK inhibitor SP600125 (final concentration of 20 μmol/L) on PMVs (D). *P < 0.05 compared with control, #P < 0.05 compared with PMV treatment. Data are means ± SE from at least three separate experiments.

Article Snippet: Phycoerythrin (PE)-conjugated anti-CD36 (clone CB38), PE-conjugated CD36 isotype control (clone G155-228), PE-cy™5-conjugated mouse anti-CD41a antibody (clone HIP8) and fluorescein isothiocyanate (FITC)-conjugated PAC-1 were from BD Biosciences/Pharmingen (San Jose, CA, USA).

Techniques: Expressing, Flow Cytometry, Binding Assay, Selection, Enzyme-linked Immunosorbent Assay, Blocking Assay, Concentration Assay

Journal: Molecular Medicine

Article Title: Oxidized Low-Density Lipoprotein-Dependent Platelet-Derived Microvesicles Trigger Procoagulant Effects and Amplify Oxidative Stress

doi: 10.2119/molmed.2011.00295

Figure Lengend Snippet: PMVs induced phosphorylation of JNK2 and its upstream activator MKK4 in CD36-positive platelets. CD36-positive and CD36-deficient platelets were incubated with PMVs and then lysed. Phosphorylated forms of JNK1/2 and MKK4 were detected by Western blot analysis for phospho-MKK4 (p-MKK4) and phospho-JNK (p-JNK1/2) and total MKK4 and JNK. (A) PMVs induced activation of JNK2 in CD36-positive but not CD36-deficient platelets. CD36-positive platelets were incubated with PMVs (30 μg/mL) for various times (B) and various concentrations of PMVs (C) for detection of phosphorylation of JNK2. Data are means ± SE of three independent experiments from different donors; *P < 0.05. □, Control; ■, PMVs.

Article Snippet: Phycoerythrin (PE)-conjugated anti-CD36 (clone CB38), PE-conjugated CD36 isotype control (clone G155-228), PE-cy™5-conjugated mouse anti-CD41a antibody (clone HIP8) and fluorescein isothiocyanate (FITC)-conjugated PAC-1 were from BD Biosciences/Pharmingen (San Jose, CA, USA).

Techniques: Incubation, Western Blot, Activation Assay

Journal: Molecular Medicine

Article Title: Oxidized Low-Density Lipoprotein-Dependent Platelet-Derived Microvesicles Trigger Procoagulant Effects and Amplify Oxidative Stress

doi: 10.2119/molmed.2011.00295

Figure Lengend Snippet: CD36-dependent platelet activation in response to PMVs. (A–C) The MFI and percentage of PAC-1, as well as platelet aggregation, were enhanced significantly after incubation of CD36-positive platelets with PMVs (30 μg/mL) for 30 min at 22°C. Data are means ± SE (n = 3). *P < 0.05 compared with control. (D–F) PAC-1 expression or platelet aggregation with CD36-deficient platelet incubation in response to PMVs. Data are means ± SE (n = 4). *P < 0.05 compared with control.

Article Snippet: Phycoerythrin (PE)-conjugated anti-CD36 (clone CB38), PE-conjugated CD36 isotype control (clone G155-228), PE-cy™5-conjugated mouse anti-CD41a antibody (clone HIP8) and fluorescein isothiocyanate (FITC)-conjugated PAC-1 were from BD Biosciences/Pharmingen (San Jose, CA, USA).

Techniques: Activation Assay, Incubation, Expressing

Journal: Molecular Medicine

Article Title: Oxidized Low-Density Lipoprotein-Dependent Platelet-Derived Microvesicles Trigger Procoagulant Effects and Amplify Oxidative Stress

doi: 10.2119/molmed.2011.00295

Figure Lengend Snippet: Identification of CD36-positive and CD36-deficient platelets by flow cytometry. MFI of binding of CD36-positive and CD36-deficient platelets with PE-conjugated anti-CD36 antibody or isotype controls.

Article Snippet: Phycoerythrin (PE)-conjugated anti-CD36 (clone CB38), PE-conjugated CD36 isotype control (clone G155-228), PE-cy™5-conjugated mouse anti-CD41a antibody (clone HIP8) and fluorescein isothiocyanate (FITC)-conjugated PAC-1 were from BD Biosciences/Pharmingen (San Jose, CA, USA).

Techniques: Flow Cytometry, Binding Assay

Journal: Molecular Medicine

Article Title: Oxidized Low-Density Lipoprotein-Dependent Platelet-Derived Microvesicles Trigger Procoagulant Effects and Amplify Oxidative Stress

doi: 10.2119/molmed.2011.00295

Figure Lengend Snippet: CD36-, PS- and JNK-dependent platelet activation by PMVs. Platelets were incubated with 2 μg/mL FA6 (A,D), 2 μg/mL mouse IgG or 20 μmol/L SP600125 (C,E) before incubation with PMVs (30 μg/mL) and analyzed by flow cytometry with FITC-labeled PAC-1. In some studies, PMVs were first incubated with 20 μg/mL annexin V to mask PS (B,D). (A–C) Histograms represent MFI and graphs; (D–E) show percentage of PAC-1–positive platelets. Data are means ± SE from at least three separate experiments. *P < 0.05 compared with control; #P < 0.05 compared with PMV treatment.

Article Snippet: Phycoerythrin (PE)-conjugated anti-CD36 (clone CB38), PE-conjugated CD36 isotype control (clone G155-228), PE-cy™5-conjugated mouse anti-CD41a antibody (clone HIP8) and fluorescein isothiocyanate (FITC)-conjugated PAC-1 were from BD Biosciences/Pharmingen (San Jose, CA, USA).

Techniques: Activation Assay, Incubation, Flow Cytometry, Labeling

Journal: Molecular Medicine

Article Title: Oxidized Low-Density Lipoprotein-Dependent Platelet-Derived Microvesicles Trigger Procoagulant Effects and Amplify Oxidative Stress

doi: 10.2119/molmed.2011.00295

Figure Lengend Snippet: (A–B) Expression of CD36 detected by flow cytometry. Preincubation of platelets with FA6, IgG, annexin V or SP600125 to examine binding of PE-conjugated CD36 antibody to platelets. (C–D) P-selection secretion determined by ELISA. The secretion of P-selection with blocking of CD36 or PS (C); the effect of JNK inhibitor SP600125 (final concentration of 20 μmol/L) on PMVs (D). *P < 0.05 compared with control, #P < 0.05 compared with PMV treatment. Data are means ± SE from at least three separate experiments.

Article Snippet: Phycoerythrin (PE)-conjugated anti-CD36 (clone CB38), PE-conjugated CD36 isotype control (clone G155-228), PE-cy™5-conjugated mouse anti-CD41a antibody (clone HIP8) and fluorescein isothiocyanate (FITC)-conjugated PAC-1 were from BD Biosciences/Pharmingen (San Jose, CA, USA).

Techniques: Expressing, Flow Cytometry, Binding Assay, Selection, Enzyme-linked Immunosorbent Assay, Blocking Assay, Concentration Assay

Journal: Molecular Medicine

Article Title: Oxidized Low-Density Lipoprotein-Dependent Platelet-Derived Microvesicles Trigger Procoagulant Effects and Amplify Oxidative Stress

doi: 10.2119/molmed.2011.00295

Figure Lengend Snippet: PMVs induced phosphorylation of JNK2 and its upstream activator MKK4 in CD36-positive platelets. CD36-positive and CD36-deficient platelets were incubated with PMVs and then lysed. Phosphorylated forms of JNK1/2 and MKK4 were detected by Western blot analysis for phospho-MKK4 (p-MKK4) and phospho-JNK (p-JNK1/2) and total MKK4 and JNK. (A) PMVs induced activation of JNK2 in CD36-positive but not CD36-deficient platelets. CD36-positive platelets were incubated with PMVs (30 μg/mL) for various times (B) and various concentrations of PMVs (C) for detection of phosphorylation of JNK2. Data are means ± SE of three independent experiments from different donors; *P < 0.05. □, Control; ■, PMVs.

Article Snippet: Phycoerythrin (PE)-conjugated anti-CD36 (clone CB38), PE-conjugated CD36 isotype control (clone G155-228), PE-cy™5-conjugated mouse anti-CD41a antibody (clone HIP8) and fluorescein isothiocyanate (FITC)-conjugated PAC-1 were from BD Biosciences/Pharmingen (San Jose, CA, USA).

Techniques: Incubation, Western Blot, Activation Assay