Journal: Nature Communications

Article Title: Binding memory of liquid molecules

doi: 10.1038/s41467-025-61630-3

Figure Lengend Snippet: Source data are provided as a Source Data file. a Schematic illustration of the 3D-SMARTER setup and the working procedure. b Trajectory of gp120-AgNP diffusing on a supported lipid bilayer. c , d 2D and 3D density maps of the residence time for the gp120-AgNP. e LDF of gp120-AgNP with a single-exponential fit. f BAF of gp120-AgNP exhibits a power law at a long-time scale. The inset displays the mean squared displacement analysis, indicating a subdiffusive exponent, \documentclass[12pt]{minimal} \usepackage{amsmath} \usepackage{wasysym} \usepackage{amsfonts} \usepackage{amssymb} \usepackage{amsbsy} \usepackage{mathrsfs} \usepackage{upgreek} \setlength{\oddsidemargin}{-69pt} \begin{document}$$\alpha=0.8460\pm 0.0002$$\end{document} α = 0.8460 ± 0.0002 . Based on Eq. , the fractal dimension of the lipid bilayer is inferred to be 1.97. Power-law fits, applied over a 0.5–8.5 s time window, quantify the scaling behavior of BAF and MSD, with exponents and fitting errors annotated in the figure. The corresponding exponents and fitting errors are indicated within the figures. g Trajectory of TATp-AgNP on a live-cell membrane. h , i 2D and 3D density maps of the residence time for TATp-AgNP. j LDF of TATp-AgNP on the live-cell membrane, with a double-exponential fit. k Power-law BAF and the MSD analysis inset, with the fractal dimension estimated to be 1.22 based on the theoretical model. Power-law fitting parameters and associated errors, extracted over the 0.25–6 s interval, are indicated in the figure.

Article Snippet: Gp120 protein (HY- P70907 , MedChemExpress) was mixed with SH-PEG-NHS (80030802E, Tanshtech) at a ratio of 1:20 and incubated overnight at 4 °C.

Techniques: Membrane

Journal: Virology

Article Title: Epitope mapping and characterization of a novel CD4-induced human monoclonal antibody capable of neutralizing primary HIV-1 strains

doi: 10.1016/S0042-6822(03)00521-X

Figure Lengend Snippet: Fig. 1. CD4 induction of the binding of the E51 antibody to HIV-1 gp120. The YU2 HIV-1 gp120 glycoprotein was captured on an ELISA plate using the D7324 antibody, which is directed against the gp120 carboxyl-termi- nus. The binding of the 17b and E51 antibodies to the captured gp120 glycoprotein was studied in the absence (open circles) or presence (filled circles) of sCD4 (8 g/ml).

Article Snippet: The ELISA plate was coated with sheep antibody D7324 (Aalto BioReagents, Dublin, Ireland), which recognizes the gp120 carboxylterminus.

Techniques: Binding Assay, Enzyme-linked Immunosorbent Assay

Journal: Journal of Neuroinflammation

Article Title: Increased neuroinflammatory and arachidonic acid cascade markers, and reduced synaptic proteins, in brain of HIV-1 transgenic rats

doi: 10.1186/1742-2094-8-101

Figure Lengend Snippet: (A) Representative immunoblot of gp120 and tat protein in HIV-1 Tg rat brain (A), detected as described in Methods . mRNA levels of brain GFAP (B) and CD11b (D) in control and HIV-1 Tg rat brain, measured using real time RT-PCR, normalized to β-globulin and relative to control level (calibrator) using the ΔΔC T method. Representative immunoblots of (C) GFAP and (E) CD11b protein in control and HIV-1 Tg rat brain. Bar graphs are ratios of optical densities of individual protein bands to β-actin, expressed as percent of control. Data represent mean ± SEM, statistical significance: **p < 0.01, ***p < 0.001 as determined by unpaired t-test.

Article Snippet: Cytosolic protein blots were incubated overnight in Tris-buffered-saline containing 5% nonfat dried milk and 0.1% Tween-20, with specific primary antibodies for proinflammatory cytokines: IL-1β (1:500), TNFα (1:500); astrocytes: glial fibrillary acidic protein (GFAP) (1:1000); CD11b (1:1000); AA cascade proteins: cPLA 2 -IVA, sPLA 2 -IIA, iPLA 2 -VIA, COX-1 (1:1000), COX-2 (1:1000), cytochrome p450 epoxygenase, TXS, 5-, 12-, 15-LOX, mPGES, cPGES (1:1000); gp120 (1:100); tat (1:100); drebrin (1:1000), BDNF (1:1000) (Santa Cruz, Santa Cruz, CA);); neurofilament-L (1:500) (Cell Signaling Technology, Danvers, MA) and β-actin (1:10,000) (Sigma Aldrich, St. Louis, MO).

Techniques: Western Blot, Control, Quantitative RT-PCR

Journal: Nanomaterials

Article Title: Calcium Phosphate Nanoparticle-Based Vaccines as a Platform for Improvement of HIV-1 Env Antibody Responses by Intrastructural Help

doi: 10.3390/nano9101389

Figure Lengend Snippet: Production and characterization of soluble HIV-1 Env constructs and Env-p30-VLPs. ( A ) Overview of different membrane-embedded and soluble Env constructs. The native BG505 gp160 (left), that is composed of three gp120 (grey) – gp41 (blue) heterodimers, has been used previously to create a soluble stabilized recombinant trimer (BG505 NFL2P gp140; middle left) by truncation at amino acid 664 and introduction of both a point mutation (I559P) indicated with an asterisk (*) and a flexible linker (2xG4S). As a purification control, we produced a BG505 NFL2P gp120 monomer (middle right) by truncation of the protein downstream of the flexible linker. A recombinant VSV-G transmembrane and cytoplasmic domain (TM/CD; red) was introduced downstream of the gp41 ectodomain to create a membrane-bound form of the stabilized trimer for production of Env-VLP-p30 particles (BG505 NFL2P gp140-GTMCD; right). ( B ) Native PAGE of purified gp140 and gp120. Three gp140 molecules formed a globular trimer molecule represented by a major band at 700 kDa. The gp120 subunit was expressed as monomeric or dimeric proteins, but did not form trimers. ( C , D ) Schematic overview of the plasmids used for Env-VLP-p30 production. ( C ) BG505 NFL2P gp140-GTMCD construct. Colors are matched to their respective protein domains in A. ( D ) Hgpsyn-TTp30 construct: the nucleotide sequence for the p30 peptide was introduced between the HIV-1 p17 matrix protein and the spacer protein 1 followed by the capsid protein p24. ( E ) Reducing SDS-PAGE and Western Blot of purified soluble Env proteins and Env-VLP-p30.

Article Snippet: The blocked membranes were incubated with polyclonal goat anti-gp120 (Acris Antibodies GmbH, Herford, Germany) and horseradish peroxidase-coupled secondary anti-goat IgG antibody (Dianova, Hamburg, Germany) with multiple washing steps in between.

Techniques: Construct, Membrane, Recombinant, Mutagenesis, Purification, Control, Produced, Clear Native PAGE, Sequencing, SDS Page, Western Blot

Journal: Journal of Virology

Article Title: A Yeast Glycoprotein Shows High-Affinity Binding to the Broadly Neutralizing Human Immunodeficiency Virus Antibody 2G12 and Inhibits gp120 Interactions with 2G12 and DC-SIGN

doi: 10.1128/jvi.02537-08

Figure Lengend Snippet: FIG. 2. 2G12 detection of a yeast protein in pmr1-containing mu- tants. (A) Equal amounts of yeast cell lysates from single-mutant and WT yeast were screened for 2G12 binding by Western blotting, with 50 ng of 293-expressed ADA gp120 as a control. (B) Cell lysate samples from the three pmr1 mnn1 clones were tested for 2G12 binding by Western blotting. (C) Culture medium samples from pmr1 and pmr1 mnn1-clone 5 (Clone 5) were blotted with 2G12 (top) and an antibody specific to the unknown protein, later identified as Pst1 (bot- tom). The migration positions of molecular mass markers, in kilodal- tons, are indicated to the left of the blots.

Article Snippet: A Yeast Glycoprotein Shows High-Affinity Binding to the Broadly Neutralizing Human Immunodeficiency Virus Antibody 2G12 and Inhibits gp120 Interactions with 2G12 and DC-SIGN Robert J. Luallen,1 Hu Fu,1 Caroline Agrawal-Gamse,2 Innocent Mboudjeka,1 Wei Huang,3 Fang-Hua Lee,2 Lai-Xi Wang,3 Robert W. Doms,2 and Yu Geng1* ProSci Incorporated, 12170 Flint Place, Poway, California 920641; Department of Microbiology, University of Pennsylvania, 225 Johnson Pavilion, Philadelphia, Pennsylvania 191042; and Institute of Human Virology, Department of Biochemistry and Molecular Biology, University of Maryland School of Medicine, 725 West Lombard Street, Baltimore, Maryland 212013 Received 9 December 2008/Accepted 19 February 2009 The human immunodeficiency virus type 1 (HIV-1) envelope (Env) protein contains numerous N-linked carbohydrates that shield conserved peptide epitopes and promote trans infection by dendritic cells via binding to cell surface lectins.

Techniques: Mutagenesis, Binding Assay, Western Blot, Control, Clone Assay, Migration

Journal: Journal of Virology

Article Title: A Yeast Glycoprotein Shows High-Affinity Binding to the Broadly Neutralizing Human Immunodeficiency Virus Antibody 2G12 and Inhibits gp120 Interactions with 2G12 and DC-SIGN

doi: 10.1128/jvi.02537-08

Figure Lengend Snippet: FIG. 5. Analysis of 2G12 reactivity between Pst1 and gp120. (A) MALDI-TOF mass spectrogram of N-linked glycans released from purified TM-Pst1. (B) A comparison of purified pmr1 mnn1-Pst1, TM-Pst1, JR-FL gp120, and Yu2 gp120 is shown with 500 ng of each protein stained by Coomassie blue (top) or 250 ng detected by Western blotting with anti-Pst1 (middle) and anti-gp120 (bottom). (C) Western blot comparing 2G12 (1 g/ml) binding to purified pmr1 mnn1-Pst1, TM-Pst1, JR-FL gp120, and Yu2 gp120 loaded at 1 g (top), 0.5 g (middle), and 0.25 g (bottom). (D) Comparison of 2G12 binding to 100 ng of purified pmr1 mnn1-Pst1 (p/m-Pst1), TM-Pst1, JR-FL gp120, and Yu2 gp120 proteins by ELISA. The values are the averages standard errors of the means (error bars) of three independent experiments. The gp120 proteins were produced in 293T cells. OD, optical density.

Article Snippet: A Yeast Glycoprotein Shows High-Affinity Binding to the Broadly Neutralizing Human Immunodeficiency Virus Antibody 2G12 and Inhibits gp120 Interactions with 2G12 and DC-SIGN Robert J. Luallen,1 Hu Fu,1 Caroline Agrawal-Gamse,2 Innocent Mboudjeka,1 Wei Huang,3 Fang-Hua Lee,2 Lai-Xi Wang,3 Robert W. Doms,2 and Yu Geng1* ProSci Incorporated, 12170 Flint Place, Poway, California 920641; Department of Microbiology, University of Pennsylvania, 225 Johnson Pavilion, Philadelphia, Pennsylvania 191042; and Institute of Human Virology, Department of Biochemistry and Molecular Biology, University of Maryland School of Medicine, 725 West Lombard Street, Baltimore, Maryland 212013 Received 9 December 2008/Accepted 19 February 2009 The human immunodeficiency virus type 1 (HIV-1) envelope (Env) protein contains numerous N-linked carbohydrates that shield conserved peptide epitopes and promote trans infection by dendritic cells via binding to cell surface lectins.

Techniques: Comparison, Staining, Western Blot, Binding Assay, Enzyme-linked Immunosorbent Assay, Produced

Journal: Journal of Virology

Article Title: A Yeast Glycoprotein Shows High-Affinity Binding to the Broadly Neutralizing Human Immunodeficiency Virus Antibody 2G12 and Inhibits gp120 Interactions with 2G12 and DC-SIGN

doi: 10.1128/jvi.02537-08

Figure Lengend Snippet: FIG. 6. TM-Pst1 binding to 2G12 by SPR. (A) Real-time SPR RU of purified TM-Pst1 (left) and Yu2 gp120 (right) from 12.5 nM to 800 nM on a 2G12-immobilized sensor chip. (B) Steady-state fitting of TM-Pst1 (left) and Yu2 gp120 (right) with the indicated KD values.

Article Snippet: A Yeast Glycoprotein Shows High-Affinity Binding to the Broadly Neutralizing Human Immunodeficiency Virus Antibody 2G12 and Inhibits gp120 Interactions with 2G12 and DC-SIGN Robert J. Luallen,1 Hu Fu,1 Caroline Agrawal-Gamse,2 Innocent Mboudjeka,1 Wei Huang,3 Fang-Hua Lee,2 Lai-Xi Wang,3 Robert W. Doms,2 and Yu Geng1* ProSci Incorporated, 12170 Flint Place, Poway, California 920641; Department of Microbiology, University of Pennsylvania, 225 Johnson Pavilion, Philadelphia, Pennsylvania 191042; and Institute of Human Virology, Department of Biochemistry and Molecular Biology, University of Maryland School of Medicine, 725 West Lombard Street, Baltimore, Maryland 212013 Received 9 December 2008/Accepted 19 February 2009 The human immunodeficiency virus type 1 (HIV-1) envelope (Env) protein contains numerous N-linked carbohydrates that shield conserved peptide epitopes and promote trans infection by dendritic cells via binding to cell surface lectins.

Techniques: Binding Assay

Journal: Journal of Virology

Article Title: A Yeast Glycoprotein Shows High-Affinity Binding to the Broadly Neutralizing Human Immunodeficiency Virus Antibody 2G12 and Inhibits gp120 Interactions with 2G12 and DC-SIGN

doi: 10.1128/jvi.02537-08

Figure Lengend Snippet: FIG. 7. TM-Pst1 inhibition of 2G12-gp120 interaction. (A) The percent 2G12 binding to 100 ng of immobilized JR-FL gp120 was analyzed by ELISA in the presence of TM-Pst1, Pst1-Endo H, Yu2 gp120, and JR-FL gp120. There was no preincubation of inhibitors with 2G12, and 100% represents 2G12 binding in the absence of inhibitor. The values are the averages standard errors of the means (error bars) of three independent experiments. Molar IC50s were cal- culated by nonlinear regression using GraphPad Prism 5.0, using the apparent molecular masses of TM-Pst1, Yu2, and JR-FL gp120 at 100 kDa (Fig. 5B). (B) (Top) Neutralization activity of MAb 2G12 against HxB-pseudotyped virus in the presence of Pst1-Endo H and TM-Pst1, with MAb 2F5 as a control. The results are representative of three separate experiments. (Bottom) Neutralization activity of 2G12 as- sessed against SF162-pseudotyped virus in the presence of Pst1-Endo H and TM-Pst1. The results are representative of two separate exper- iments.

Article Snippet: A Yeast Glycoprotein Shows High-Affinity Binding to the Broadly Neutralizing Human Immunodeficiency Virus Antibody 2G12 and Inhibits gp120 Interactions with 2G12 and DC-SIGN Robert J. Luallen,1 Hu Fu,1 Caroline Agrawal-Gamse,2 Innocent Mboudjeka,1 Wei Huang,3 Fang-Hua Lee,2 Lai-Xi Wang,3 Robert W. Doms,2 and Yu Geng1* ProSci Incorporated, 12170 Flint Place, Poway, California 920641; Department of Microbiology, University of Pennsylvania, 225 Johnson Pavilion, Philadelphia, Pennsylvania 191042; and Institute of Human Virology, Department of Biochemistry and Molecular Biology, University of Maryland School of Medicine, 725 West Lombard Street, Baltimore, Maryland 212013 Received 9 December 2008/Accepted 19 February 2009 The human immunodeficiency virus type 1 (HIV-1) envelope (Env) protein contains numerous N-linked carbohydrates that shield conserved peptide epitopes and promote trans infection by dendritic cells via binding to cell surface lectins.

Techniques: Inhibition, Binding Assay, Enzyme-linked Immunosorbent Assay, Neutralization, Activity Assay, Virus, Control

Journal: Journal of Virology

Article Title: A Yeast Glycoprotein Shows High-Affinity Binding to the Broadly Neutralizing Human Immunodeficiency Virus Antibody 2G12 and Inhibits gp120 Interactions with 2G12 and DC-SIGN

doi: 10.1128/jvi.02537-08

Figure Lengend Snippet: FIG. 8. TM-Pst1 inhibition of DC-SIGN–gp120 interaction. (A) Graded concentrations of TM-Pst1, mannan, and BSA were eval- uated for direct inhibition of soluble JR-FL gp120 binding to immo- bilized DC-SIGN by ELISA. The results are shown relative to 100% gp120 binding, calculated in the absence of inhibitor. The values are the averages standard errors of the means (error bars) of three independent experiments for Pst1 and mannan and two for BSA. IC50s were calculated by nonlinear regression using GraphPad Prism 5. (B) (Top) Inhibition of JRFL gp120 (1 g) binding to THP–DC-SIGN by 10 g of mannan, BSA, or TM-Pst1. (Bottom) Dose-dependent inhibition of JR-FL gp120 (1 g) binding to THP–DC-SIGN cells by TM-Pst1. As controls, cells were incubated with only antibodies (no gp120) or gp120 without inhibitors (gp120). The results are represen- tative of two independent experiments.

Article Snippet: A Yeast Glycoprotein Shows High-Affinity Binding to the Broadly Neutralizing Human Immunodeficiency Virus Antibody 2G12 and Inhibits gp120 Interactions with 2G12 and DC-SIGN Robert J. Luallen,1 Hu Fu,1 Caroline Agrawal-Gamse,2 Innocent Mboudjeka,1 Wei Huang,3 Fang-Hua Lee,2 Lai-Xi Wang,3 Robert W. Doms,2 and Yu Geng1* ProSci Incorporated, 12170 Flint Place, Poway, California 920641; Department of Microbiology, University of Pennsylvania, 225 Johnson Pavilion, Philadelphia, Pennsylvania 191042; and Institute of Human Virology, Department of Biochemistry and Molecular Biology, University of Maryland School of Medicine, 725 West Lombard Street, Baltimore, Maryland 212013 Received 9 December 2008/Accepted 19 February 2009 The human immunodeficiency virus type 1 (HIV-1) envelope (Env) protein contains numerous N-linked carbohydrates that shield conserved peptide epitopes and promote trans infection by dendritic cells via binding to cell surface lectins.

Techniques: Inhibition, Binding Assay, Enzyme-linked Immunosorbent Assay, Incubation

Journal: International journal of molecular sciences

Article Title: KYNA Ameliorates Glutamate Toxicity of HAND by Enhancing Glutamate Uptake in A2 Astrocytes.

doi: 10.3390/ijms25084286

Figure Lengend Snippet: Figure 1. The effect of HIV-1 gp120 on the glutamate uptake capacity of primary astrocytes. (A) Viability of primary astrocytes by CCK8 measurement of incremental concentrations of L- glutamate. (B) Glutamate concentration in the supernatant of gp120-treated primary astrocytes. (C) Influence of gp120 on glutamate uptake capacity of primary astrocytes. (D) Glutamate content in the cerebrospinal fluid of WT mice and gp120 tgm mice. (E) gp120 inhibits EAAT2 protein expression in primary astrocytes. ** p < 0.01, and *** p < 0.001.

Article Snippet: Cultured astrocytes were treated with 0.01 μg/mL of HIV-1 gp120 [HIV-1/Clade B/C (CN54), China] for 12 h, followed by the administration of 25 μM KYNA (Sigma, St. Louis, MO, USA) for 12 h. In addition, astrocytes were pretreated with 10 nM methyllycaconitine (MLA, MCE, Monmouth Junction, NJ, USA), 500 nM PNU-120596 (MCE, NJ, USA) or 5 μM Bay11-7085 (GLPBIO, Montclair, CA, USA) for 1 h, respectively.

Techniques: Concentration Assay, Expressing

Journal: International journal of molecular sciences

Article Title: KYNA Ameliorates Glutamate Toxicity of HAND by Enhancing Glutamate Uptake in A2 Astrocytes.

doi: 10.3390/ijms25084286

Figure Lengend Snippet: Figure 2. The effect of HIV−1 gp120 on mouse brain A2 astrocytes. Spectrum of mRNA expression of A2−specific genes (A) and trophic factors (C) in mouse brain parenchyma. (B) C3 protein levels in brain parenchyma of WT mice and gp120 tgm mice. (D) Expression levels of C3 protein in gp120−treated primary astrocytes. ** p < 0.01, and *** p < 0.001.

Article Snippet: Cultured astrocytes were treated with 0.01 μg/mL of HIV-1 gp120 [HIV-1/Clade B/C (CN54), China] for 12 h, followed by the administration of 25 μM KYNA (Sigma, St. Louis, MO, USA) for 12 h. In addition, astrocytes were pretreated with 10 nM methyllycaconitine (MLA, MCE, Monmouth Junction, NJ, USA), 500 nM PNU-120596 (MCE, NJ, USA) or 5 μM Bay11-7085 (GLPBIO, Montclair, CA, USA) for 1 h, respectively.

Techniques: Expressing

Journal: International journal of molecular sciences

Article Title: KYNA Ameliorates Glutamate Toxicity of HAND by Enhancing Glutamate Uptake in A2 Astrocytes.

doi: 10.3390/ijms25084286

Figure Lengend Snippet: Figure 3. KYNA promotes A2 astrocyte formation. (A) The mRNA expression profiles of A2−specific genes in KYNA and gp120−treated primary astrocytes. (B) Trophic factor content in the supernatant of primary astrocytes. (C) Quantification of C3 protein levels in primary astrocytes by Western blot. (D) Immunofluorescence images showing the expression of C3 in primary astrocytes (scale bar: 50 µm), and quantified using ImageJ software (ImageJ 1.54f). * p < 0.05, ** p < 0.01, and *** p < 0.001.

Article Snippet: Cultured astrocytes were treated with 0.01 μg/mL of HIV-1 gp120 [HIV-1/Clade B/C (CN54), China] for 12 h, followed by the administration of 25 μM KYNA (Sigma, St. Louis, MO, USA) for 12 h. In addition, astrocytes were pretreated with 10 nM methyllycaconitine (MLA, MCE, Monmouth Junction, NJ, USA), 500 nM PNU-120596 (MCE, NJ, USA) or 5 μM Bay11-7085 (GLPBIO, Montclair, CA, USA) for 1 h, respectively.

Techniques: Expressing, Western Blot, Immunofluorescence, Software

Journal: International journal of molecular sciences

Article Title: KYNA Ameliorates Glutamate Toxicity of HAND by Enhancing Glutamate Uptake in A2 Astrocytes.

doi: 10.3390/ijms25084286

Figure Lengend Snippet: Figure 4. Effect of KYNA on glutamate uptake capacity of primary astrocytes. (A) Glutamate content in the supernatant of KYNA−treated primary astrocytes. (B) Expression of EAAT2 in KYNA and gp120−treated primary astrocytes was assessed by Western blot. (C) Immunofluorescence images showing the expression of EAAT2 in primary astrocytes (scale bar: 50 µm), and quantitatively analyzed using ImageJ software (ImageJ 1.54f). ** p < 0.01, and ***p < 0.001.

Article Snippet: Cultured astrocytes were treated with 0.01 μg/mL of HIV-1 gp120 [HIV-1/Clade B/C (CN54), China] for 12 h, followed by the administration of 25 μM KYNA (Sigma, St. Louis, MO, USA) for 12 h. In addition, astrocytes were pretreated with 10 nM methyllycaconitine (MLA, MCE, Monmouth Junction, NJ, USA), 500 nM PNU-120596 (MCE, NJ, USA) or 5 μM Bay11-7085 (GLPBIO, Montclair, CA, USA) for 1 h, respectively.

Techniques: Expressing, Western Blot, Immunofluorescence, Software

Journal: International journal of molecular sciences

Article Title: KYNA Ameliorates Glutamate Toxicity of HAND by Enhancing Glutamate Uptake in A2 Astrocytes.

doi: 10.3390/ijms25084286

Figure Lengend Snippet: Figure 5. Effects of α7nAChR antagonist (MLA) and agonist (PNU−282987) on A2 astro- cyte formation. (A,D) The mRNA expression profiles of A2−specific genes in the MLA−and PNU−282987−treated primary astrocytes. (B) Trophic factor content in the supernatant of primary astrocytes. Expression levels of GFAP (C,G) and C3 (F,H) in MLA−and PNU−282987−treated primary astrocytes. (E) mRNA expression profiles of trophic factors in PNU−282987−treated A2 astrocytes. (I,J) Glutamate content in the supernatant of MLA and PNU−282987−treated primary astrocytes. * represents comparison with control group. NS means p > 0.05, * p < 0.05, ** p < 0.01, and *** p < 0.001. $ represents comparison with HIV-1 gp120 group. $ represents p < 0.05, $$$ represents p < 0.001.

Article Snippet: Cultured astrocytes were treated with 0.01 μg/mL of HIV-1 gp120 [HIV-1/Clade B/C (CN54), China] for 12 h, followed by the administration of 25 μM KYNA (Sigma, St. Louis, MO, USA) for 12 h. In addition, astrocytes were pretreated with 10 nM methyllycaconitine (MLA, MCE, Monmouth Junction, NJ, USA), 500 nM PNU-120596 (MCE, NJ, USA) or 5 μM Bay11-7085 (GLPBIO, Montclair, CA, USA) for 1 h, respectively.

Techniques: Expressing, Comparison, Control

Journal: International journal of molecular sciences

Article Title: KYNA Ameliorates Glutamate Toxicity of HAND by Enhancing Glutamate Uptake in A2 Astrocytes.

doi: 10.3390/ijms25084286

Figure Lengend Snippet: Figure 6. The effects of α7nAChR antagonist (MLA) and agonist (PNU−282987) on the glutamate uptake capacity of A2 astrocytes. (A) Levels of α7nAChR in gp120−treated primary astrocytes. The levels of MLA−and PNU−282987−treated primary astrocytes α7nAChR proteins (B,D), and EAAT2 proteins (C,E). Immunofluorescence showed the expression of EAAT2 protein in MLA−(F) and PNU−282987−(G) treated primary astrocytes (scale bar: 50 µm), and quantitative analysis was performed using ImageJ software (ImageJ 1.54f). * p < 0.05, ** p < 0.01, and *** p < 0.001.

Article Snippet: Cultured astrocytes were treated with 0.01 μg/mL of HIV-1 gp120 [HIV-1/Clade B/C (CN54), China] for 12 h, followed by the administration of 25 μM KYNA (Sigma, St. Louis, MO, USA) for 12 h. In addition, astrocytes were pretreated with 10 nM methyllycaconitine (MLA, MCE, Monmouth Junction, NJ, USA), 500 nM PNU-120596 (MCE, NJ, USA) or 5 μM Bay11-7085 (GLPBIO, Montclair, CA, USA) for 1 h, respectively.

Techniques: Immunofluorescence, Expressing, Software

Journal: International journal of molecular sciences

Article Title: KYNA Ameliorates Glutamate Toxicity of HAND by Enhancing Glutamate Uptake in A2 Astrocytes.

doi: 10.3390/ijms25084286

Figure Lengend Snippet: Figure 8. Effect of NF−κB antagonist (Bay11−7085) on glutamate uptake capacity of A2 astrocytes. (A) Immunofluorescence showing the expression of NF−κB subunit pp65 in primary astrocytes after Bay11−7085 treatment (scale bar: 10 µm). (B) Protein expression of pp65 in gp120−treated primary astrocytes. Protein levels of α7nAChR (C), NF−κB (D), and EAAT2 (E) were measured by Western blot in Bay11−7085−treated astrocytes. * p < 0.05, ** p < 0.01, and *** p < 0.001.

Article Snippet: Cultured astrocytes were treated with 0.01 μg/mL of HIV-1 gp120 [HIV-1/Clade B/C (CN54), China] for 12 h, followed by the administration of 25 μM KYNA (Sigma, St. Louis, MO, USA) for 12 h. In addition, astrocytes were pretreated with 10 nM methyllycaconitine (MLA, MCE, Monmouth Junction, NJ, USA), 500 nM PNU-120596 (MCE, NJ, USA) or 5 μM Bay11-7085 (GLPBIO, Montclair, CA, USA) for 1 h, respectively.

Techniques: Immunofluorescence, Expressing, Western Blot

Journal: International journal of molecular sciences

Article Title: KYNA Ameliorates Glutamate Toxicity of HAND by Enhancing Glutamate Uptake in A2 Astrocytes.

doi: 10.3390/ijms25084286

Figure Lengend Snippet: Figure 9. The effects of intraperitoneal injection of KYN and PROB on the metabolism of the KP pathway in mice. (A) KYN content in blood of WT and gp120 tgm mice. KYN (B), KYNA (C), and QUIN (D) levels in mouse cerebrospinal fluid were measured by ELISA. (E) KYNA/QUIN ratio in mouse cerebrospinal fluid. * p < 0.05, ** p < 0.01, and *** p < 0.001.

Article Snippet: Cultured astrocytes were treated with 0.01 μg/mL of HIV-1 gp120 [HIV-1/Clade B/C (CN54), China] for 12 h, followed by the administration of 25 μM KYNA (Sigma, St. Louis, MO, USA) for 12 h. In addition, astrocytes were pretreated with 10 nM methyllycaconitine (MLA, MCE, Monmouth Junction, NJ, USA), 500 nM PNU-120596 (MCE, NJ, USA) or 5 μM Bay11-7085 (GLPBIO, Montclair, CA, USA) for 1 h, respectively.

Techniques: Injection, Enzyme-linked Immunosorbent Assay

Journal: International journal of molecular sciences

Article Title: KYNA Ameliorates Glutamate Toxicity of HAND by Enhancing Glutamate Uptake in A2 Astrocytes.

doi: 10.3390/ijms25084286

Figure Lengend Snippet: Figure 11. Effects of intraperitoneal injection of KYN and PROB on mouse brain A2 astrocytes. (A) Representative images of immunohistochemical staining of GFAP on cortical and hippocam- pal brain sections from gp120+/α7nAChR−/−mice and gp120 tgm mice (scale bar: 50 µM). (B) A1 astrocyte marker C3 protein levels in mouse cerebrospinal fluid. (C) ELISA detected trophic factor levels in mouse cerebrospinal fluid. The mRNA expression profiles of A2−specific genes (D) and trophic factor genes (E) in mouse brain parenchyma. * p < 0.05, ** p < 0.01, and *** p < 0.001.

Article Snippet: Cultured astrocytes were treated with 0.01 μg/mL of HIV-1 gp120 [HIV-1/Clade B/C (CN54), China] for 12 h, followed by the administration of 25 μM KYNA (Sigma, St. Louis, MO, USA) for 12 h. In addition, astrocytes were pretreated with 10 nM methyllycaconitine (MLA, MCE, Monmouth Junction, NJ, USA), 500 nM PNU-120596 (MCE, NJ, USA) or 5 μM Bay11-7085 (GLPBIO, Montclair, CA, USA) for 1 h, respectively.

Techniques: Injection, Immunohistochemical staining, Staining, Marker, Enzyme-linked Immunosorbent Assay, Expressing

Journal: International journal of molecular sciences

Article Title: KYNA Ameliorates Glutamate Toxicity of HAND by Enhancing Glutamate Uptake in A2 Astrocytes.

doi: 10.3390/ijms25084286

Figure Lengend Snippet: Figure 12. Effects of intraperitoneal injection of KYN and PROB on glutamate in mouse brain. (A) Immunofluorescence showing pp65 expression in the hippocampus and cortex of WT and gp120 tgm mice (scale bar: 50 µm). (B) Glutamate content in the brain of WT and gp120 tgm mice. (C) Immunohistochemistry showing the expression of EAAT2 in the hippocampus and cortex of WT and gp120 tgm mice (scale bar: 50 µm), and quantitative analysis using ImageJ software (ImageJ 1.54f). * p < 0.05, ** p < 0.01, and *** p < 0.001.

Article Snippet: Cultured astrocytes were treated with 0.01 μg/mL of HIV-1 gp120 [HIV-1/Clade B/C (CN54), China] for 12 h, followed by the administration of 25 μM KYNA (Sigma, St. Louis, MO, USA) for 12 h. In addition, astrocytes were pretreated with 10 nM methyllycaconitine (MLA, MCE, Monmouth Junction, NJ, USA), 500 nM PNU-120596 (MCE, NJ, USA) or 5 μM Bay11-7085 (GLPBIO, Montclair, CA, USA) for 1 h, respectively.

Techniques: Injection, Immunofluorescence, Expressing, Immunohistochemistry, Software

Journal: International journal of molecular sciences

Article Title: KYNA Ameliorates Glutamate Toxicity of HAND by Enhancing Glutamate Uptake in A2 Astrocytes.

doi: 10.3390/ijms25084286

Figure Lengend Snippet: Figure 13. The effects of intraperitoneal injection of KYN and PROB on memory, cognition and mood in mice. (A) Latency of hidden platform training for 5 consecutive days in each group of mice. (B) La- tency of hidden platform training for 5 consecutive days in gp120 tgm mice intraperitoneally injected with KYN and PROB and in gp120 tgm mice intraperitoneally injected with PBS. (C) Representative trajectories of mice in the spatial exploration test. (D) The number of times mice traversed the plat- form in the spatial exploration experiment. Time spent by mice exploring the center region (E) and four quadrants (F) of the water maze device. The number of times mice traversed the center region of the open field device (G), the time spent (J), and the distance moved (K). Representative trajectories (I) and total distance moved (H) by mice in the open field experiment. * p < 0.05, ** p < 0.01, and *** p < 0.001.

Article Snippet: Cultured astrocytes were treated with 0.01 μg/mL of HIV-1 gp120 [HIV-1/Clade B/C (CN54), China] for 12 h, followed by the administration of 25 μM KYNA (Sigma, St. Louis, MO, USA) for 12 h. In addition, astrocytes were pretreated with 10 nM methyllycaconitine (MLA, MCE, Monmouth Junction, NJ, USA), 500 nM PNU-120596 (MCE, NJ, USA) or 5 μM Bay11-7085 (GLPBIO, Montclair, CA, USA) for 1 h, respectively.

Techniques: Injection