scavenger receptors expression  (Human Protein Atlas)

Journal: Blood

Article Title: Oligo-guanosine nucleotide induces neuropilin-1 internalization in endothelial cells and inhibits angiogenesis

doi: 10.1182/blood-2010-01-265801

Figure Lengend Snippet: Selective reduction of NRP1 by G18 is temperature-dependent. (A) Effects of G18 on levels of cell-surface NRP1, NRP2, VEGFR-2, VEGFR1, gp130, CD31, and SREC-I detected by flow cytometry. HUVECs were incubated at 37°C for 1 hour with or without G18 (16 μg/mL). Shaded graphs reflect control staining. (B) Temperature- and time-dependent reduction of cell-surface NRP1 by G18. HUVECs were incubated with G18 (16 μg/mL) at 4° or 37°C for 5, 15, 30, and 60 minutes. The results reflect the relative mean fluorescence intensities of NRP1 under the conditions of testing. (C) Levels of cell-surface NRP1 detected by flow cytometry on HUVECs incubated with sG18 (0, 1, 2, 4, 8, or 16 μg/mL) in the presence of 1% FBS or with sG18 (0, 4, 8, 16, 32, or 64 μg/mL) in the presence of 95% FBS.

Article Snippet: Reagents and cytokines Recombinant human VEGF 165 , human interleukin-6 (IL-6), soluble IL-6 receptor (sIL-6R), chimeric rat NRP1/Fc, NRP2/Fc, human Sema3A/Fc, and scavenger receptor expressed by endothelial cells I (SREC-I)/Fc were from R&D Systems.

Techniques: Flow Cytometry, Incubation, Staining, Fluorescence

Journal: Blood

Article Title: Oligo-guanosine nucleotide induces neuropilin-1 internalization in endothelial cells and inhibits angiogenesis

doi: 10.1182/blood-2010-01-265801

Figure Lengend Snippet: Characterization and functional consequences of G18 binding to NRP1 and SREC-I. (A) Analysis of biotin-G18 binding to NRP1-Fc and NRP2-Fc. Top panel, binding of biotin-G18 to BSA-, IgG1-, NRP1-, or NRP2-coated wells detected by ELISA-based measurement of streptavidin-HRP. Bottom panel, detection of coated proteins by anti-IgG1 antibody detected by ELISA. The results reflect the means ± SD of triplicate wells. (B) Binding of biotin-G18 to NRP1. Biotin-A18, -T18, -G18, or -C18 (0.25, 1, 4, 16 μg/mL) was added to NRP1-coated wells. Bound biotin-olionucleotides were detected by ELISA-based measurement of streptavidin-HRP. The results reflect the means ± SD of 3 experiments. (C) Biotin-G18 binding to NRP1 is selectively blocked by nonbiotin-G18. Biotin-G18 (1 μg/mL) was added to NRP1-coated wells in the presence of G18, A18, T18, or C18 (100 μg/mL). (D) Characterization of NRP1 binding to immobilized G18. Plasmon resonance (Biacore)-generated sensorgrams showing a kinetic analysis of NRP1 binding to G18. Biotin-G18 was immobilized onto the sensor chip. NRP1/Fc (2.5, 5, 7.5, 10, or 20nM) was passed over the sensor surface. Representative results from 4 independent experiments are shown. (E) Binding of biotin-G18 to SREC-I. Biotin-A18, -T18, -G18, or -C18 (0.25, 1, 4, or 16 μg/mL) was added to SREC-I–coated wells. Bound biotin-olionucleotides were detected by ELISA-based measurement of streptavidin-HRP. The results reflect the means ± SD of 3 experiments. (F) G18 bridges NRP1 to SREC-I. His-tagged NRP1/Fc (1 μg/mL) was added to SREC-I/Fc-coated wells in the presence of A18, T18, G18, or C18 (0, 0.25, 1, 4, or 16 μg/mL). Bound His-tagged NRP1/Fc was detected by HRP-conjugated anti-His Ab. The results reflect the means ± SD of 3 experiments. (G) G18 induces the coordinate internalization of NRP1 and SREC-I. HUVECs were incubated with or without sG18 (16 μg/mL at 37°C for 1 hour). After fixation and permeabilization, cells were stained with anti-NRP1 mAb (green), anti-SREC-I Ab (red), and DAPI (blue) and were examined by confocal microscopy. Top panels are from cells incubated in medium alone; bottom panels are from cells incubated with sG18. Scale bar, 20 μm.

Article Snippet: Reagents and cytokines Recombinant human VEGF 165 , human interleukin-6 (IL-6), soluble IL-6 receptor (sIL-6R), chimeric rat NRP1/Fc, NRP2/Fc, human Sema3A/Fc, and scavenger receptor expressed by endothelial cells I (SREC-I)/Fc were from R&D Systems.

Techniques: Functional Assay, Binding Assay, Enzyme-linked Immunosorbent Assay, Generated, Incubation, Staining, Confocal Microscopy