Journal: PLoS ONE

Article Title: Reduction of the HIV Protease Inhibitor-Induced ER Stress and Inflammatory Response by Raltegravir in Macrophages

doi: 10.1371/journal.pone.0090856

Figure Lengend Snippet: Real time PCR primers.

Article Snippet: Mouse monoclonal antibody against CD36 was from Cayman Chemical (Ann Arbor, MI).

Techniques: Real-time Polymerase Chain Reaction

Journal: PLoS ONE

Article Title: Reduction of the HIV Protease Inhibitor-Induced ER Stress and Inflammatory Response by Raltegravir in Macrophages

doi: 10.1371/journal.pone.0090856

Figure Lengend Snippet: ( A–C ) Human THP-1-derived macrophages were treated with HIV PIs (lopinavir/Ritonavir, LOPV/RITV, 25 µM) with or without raltegravir (25 µM) for 24 h. The total cellular RNA was isolated and reverse transcribed. The relative mRNA levels of CD36, SRA, and LDLR were determined by real-time PCR as described in “Methods”. The values are means ± S.E. of three independent experiments. **, p<0.01, ***, p<0.001, statistical significance relative to vehicle control. #, p<0.05, statistical significance of HIV PI+raltegravir-treated group relative to HIV PI-treated group. ( D ) Human THP-1-derived macrophages were treated with HIV PIs (lopinavir/Ritonavir, LOPV/RITV, 25 µM) with or without raltegravir (25 µM) for 24 h. The total protein lysates were prepared and subjected to Western blot analysis. Representative immunoblots for CD36, SRA and β-actin are shown. β-actin was used as the loading control for total proteins.

Article Snippet: Mouse monoclonal antibody against CD36 was from Cayman Chemical (Ann Arbor, MI).

Techniques: Derivative Assay, Isolation, Reverse Transcription, Real-time Polymerase Chain Reaction, Control, Western Blot

Journal: PLoS ONE

Article Title: Oxidation by Neutrophils-Derived HOCl Increases Immunogenicity of Proteins by Converting Them into Ligands of Several Endocytic Receptors Involved in Antigen Uptake by Dendritic Cells and Macrophages

doi: 10.1371/journal.pone.0123293

Figure Lengend Snippet: ( A ) Uptake of fluorescently-labelled proteins by CHO cells transfected with human CD36 as compared to non-transfected cells. ( B ) Binding of anti-CD36 mAb and control mouse IgA to CD36-transfected and non-transfected CHO cells, determined by cellular ELISA. ( C ) Binding of rCD36 to plate-adsorbed proteins. ( D ) Effects of 10 or 100 μg/ml of indicated, soluble ligands on rCD36 binding to plate-adsorbed OVA-Cl. ( E ) Binding of polyclonal anti-mouse CD36 Ab and control goat IgG to BM-DC, splenic DC and PEM. A representative histogram of Ab binding to BM-DC is displayed on the left graph. ( F ) Binding of anti-mouse SR-A mAb and control rat IgG2b to BM-DC, splenic DC and PEM. A representative histogram of Ab binding to BM-DC is shown on the left graph. ( G ) Binding of SR-A present in lysates of PEM to plate-adsorbed proteins. ( H ) Effects of anti-SR-A 2F8 mAb and AcLDL on the uptake of fluorescently-labelled proteins by BM-DC. Shown are results of single experiments, each representative of at least 3 similar experiments performed (A-D, G, H) or averages +SEM from 4–6 independent experiments (E, F). The data were analysed with the unpaired (A-C, G) or one-sample (H) Student’s t-test or with ANOVA, followed by the Tukey-Kramer post-test (D). *, p < 0.05; ND, not done.

Article Snippet: Rat anti-mouse scavenger receptor A (SR-A) mAb (clone 2F8) was obtained from AbD Serotec; mouse anti-mouse CD36 mAb (CRF D-2712) from Hycult Biotech; mouse IgA isotype control mAb (M18-254), rat IgG2b isotype control mAb (A95-1), rat anti-mouse CD11b mAb (M1/70) and phycoerythrin (PE)-streptavidin conjugate from BD Biosciences; rat IgG2a isotype control mAb (54447), normal goat IgG, polyclonal goat anti-mouse CD36, anti-mouse LOX-1 (lectin-type oxidised LDL receptor-1), anti-human SREC-I (scavenger receptor expressed by endothelial cells-I) Ab and PE-conjugated rat anti-mouse LOX-1 mAb (214012) from R&D Systems; polyclonal goat anti-mouse SREC-I, anti-mouse RAGE (receptor for advanced glycation end products), anti-mouse stabilin-1 and rabbit anti-mouse-stabilin-1 Ab from Santa Cruz Biotechnology; PE-conjugated donkey anti-goat IgG Ab from SouthernBiotech; rat anti-mouse CD206/mannose receptor mAb (MR5D3) and PE-conjugated goat anti-rat IgG Ab from BioLegend; horseradish peroxidase (HRP)-conjugated rabbit anti-mouse IgA, F(ab’)2 fragments of goat anti-rat IgG and donkey anti-goat IgG Ab from Rockland.

Techniques: Transfection, Binding Assay, Enzyme-linked Immunosorbent Assay