Journal: Redox Biology

Article Title: Radiosensitizing capacity of fenofibrate in glioblastoma cells depends on lipid metabolism

doi: 10.1016/j.redox.2024.103452

Figure Lengend Snippet: Fenofibrate (FF) modulates lipid droplets (LDs) in U87 and LN18 cells. A) Schematic representation of uptake and processing pathways of fatty acids (FAs) in tumor cells. B-i) Percentage of U87 and LN18 cells expressing CD36. B-ii) Mean fluorescent intensity (MFI) of CD36 expression in U87 and LN18 cells. C) U87 and LN18 cells were stained with LipidTOX™ (red) for LD, GPAT4 antibody (green) for lipid enzyme and Hoechst (blue) for nuclei. D) Representative immunoblots and bar graphs (from 3 independent experiments) of basal GPAT4 (D-i) and DGAT1 (D-ii) expression in U87 and LN18 cells. E) Representative confocal images of LDs (red) and nuclei (blue) of U87 and LN18 cells for all treatment groups. F) Graph showing numbers of small and large LDs in U87 (top) and LN18 (bottom) cells per treatment. LD quantification was performed by counting red lipid bodies using ImageJ (minimum of 50 cells per cell line). G) Representative immunoblots and bar graphs showing the expression of DGAT1 in U87 (top) and LN18 (bottom) cells after FF and RTx treatment. H) The normalized value of intracellular GPAT4 in U87 (top) and LN18 (bottom) cells in all treatment groups. Unless stated otherwise, data are shown as means ± SD of at least three independent experiments. P values were calculated using an unpaired t -test (two groups) or a two-way analysis of variance (ANOVA, more than two groups) with Tukey's correction. ∗p < 0.0332, ∗∗p < 0.0021, ∗∗∗p < 0.0002 and ∗∗∗∗p < 0.0001.

Article Snippet: Cells (0.25x10 6 ) were trypsinized, washed twice with ice-cold flow cytometry buffer (phosphate-buffered saline (PBS; Life Technologies)), containing 10 % v/v FBS (Sigma-Aldrich) and incubated with FITC-conjugated CD36 monoclonal antibody (mAb, AC106, 2 μg/mL, Miltenyi Biotec), and membrane Hsp70 mAb (cmHsp70.1, 20 μg/mL, multimmune GmbH) for 30 min in the dark on ice.

Techniques: Expressing, Staining, Western Blot

Journal: Cancers

Article Title: Murlentamab, a Low Fucosylated Anti-Müllerian Hormone Type II Receptor (AMHRII) Antibody, Exhibits Anti-Tumor Activity through Tumor-Associated Macrophage Reprogrammation and T Cell Activation

doi: 10.3390/cancers13081845

Figure Lengend Snippet: Flow cytometry antibodies used.

Article Snippet: CD36-PEVio770 , Miltenyi , 130-110-742 , REA770 , .

Techniques: Flow Cytometry, In Vivo, In Vitro

Journal: Cancers

Article Title: Murlentamab, a Low Fucosylated Anti-Müllerian Hormone Type II Receptor (AMHRII) Antibody, Exhibits Anti-Tumor Activity through Tumor-Associated Macrophage Reprogrammation and T Cell Activation

doi: 10.3390/cancers13081845

Figure Lengend Snippet: Murlentamab opsonization of SKOV3-R2 + orients naïve macrophages and reprograms TAMs towards an M1-like profile. SKOV3-R2 + ovarian tumor cells were labeled with different 3C23K antibodies (3C23K-FcKO control, 3C23K-CHO normally fucosylated or murlentamab the low fucosylated form) and cultured in the presence of human monocyte-derived macrophages from healthy donors unstimulated (M0) or stimulated with M-CSF and IL-10 (TAMs). ( A ) The proportion of macrophages expressing M1/M2 membrane markers (CD32, CD64, CD80, TLR2, CD163, CD36 and CD206) was determined by flow cytometry after three days of co-culture with SKOV3-R2 + cells. ( B ) The release of cytokines (IL1β, IL12, TNFα, IL6, IFNγ, IL10) and chemokines (CCL2, CCL4, CCL5, CXCL9 and CXCL10) in the culture medium was determined by AlphaLISA after three days of co-culture with SKOV3-R2 + cells. Data shown (boxplots) are the results from three different experiments (performed with three different healthy donors). * p < 0.05; ** p < 0.01; *** p < 0.001; **** p < 0.0001. p values were determined using one-way ANOVA analysis followed by Tukey’s multiple comparisons test.

Article Snippet: CD36-PEVio770 , Miltenyi , 130-110-742 , REA770 , .

Techniques: Labeling, Cell Culture, Derivative Assay, Expressing, Membrane, Flow Cytometry, Co-Culture Assay

Journal: Frontiers in Immunology

Article Title: Crystalline Silica Impairs Efferocytosis Abilities of Human and Mouse Macrophages: Implication for Silica-Associated Systemic Sclerosis

doi: 10.3389/fimmu.2020.00219

Figure Lengend Snippet: Impairment of efferocytosis by silica in MDM requires SR-B1 expression. M0 MDM from the same healthy donors, were transfected with siRNA Ct or siRNA for SR-B1 for 24 h. Percentages of SR-B1 positive MDM (A) and EI of MDM from the same 4 healthy donors (B) , untreated or treated to SiO 2 and then exposed to apoptotic Jurkat (apoJ) or live Jurkat (J) cells for 90 min, were both determined by flow cytometry (Experiment on MDM from 4 different healthy donors). * p < 0.05; ** p < 0.01; ns, not significant.

Article Snippet: MDM were transfected using Lipofectamine RNAiMax reagent (Invitrogen) with siRNA at 5 pmol for 24 h. Silencing efficiency of siRNA SR-B1 was analyzed at the protein level using the anti-SR-B1-APC antibody (Miltenyi Biotec, SAS) by flow cytometry on a LSR II cytometer.

Techniques: Expressing, Transfection, Flow Cytometry

Journal: Frontiers in Immunology

Article Title: Crystalline Silica Impairs Efferocytosis Abilities of Human and Mouse Macrophages: Implication for Silica-Associated Systemic Sclerosis

doi: 10.3389/fimmu.2020.00219

Figure Lengend Snippet: RhoA/ROCK pathway inhibition enhances efferocytosis capacities of silica-exposed MDM and this effect is not due to a down expression of SR-B1 after RhoA/ROCK inhibition. (A,B) EI of MDM pre-treated or not with the ROCK inhibitors (A) Y27632 or (B) fasudil at 20 μM and treated with 25 μg/cm 2 of SiO 2 for 4 h and then exposed to apoptotic Jurkat cells for 90 min (Experiment on MDM from at least 5 different healthy donors). (C,D) Expression of SR-B1 expressed as (C) ratio of MFI and (D) percentage of positive cells of MDM treated or not with the ROCK inhibitor Y27632 and Fasudil at 20 μM for 4 h analyzed by flow cytometry (Experiment on MDM from 2 to 5 different healthy donors). * p < 0.05; ** p < 0.01; ns, not significant.

Article Snippet: MDM were transfected using Lipofectamine RNAiMax reagent (Invitrogen) with siRNA at 5 pmol for 24 h. Silencing efficiency of siRNA SR-B1 was analyzed at the protein level using the anti-SR-B1-APC antibody (Miltenyi Biotec, SAS) by flow cytometry on a LSR II cytometer.

Techniques: Inhibition, Expressing, Flow Cytometry