Journal: Cell host & microbe

Article Title: NON-NEUTRALIZING ANTIBODIES FROM A MARBURG INFECTION SURVIVOR MEDIATE PROTECTION BY FC-EFFECTOR FUNCTIONS AND ENHANCING EFFICACY OF OTHER ANTIBODIES

doi: 10.1016/j.chom.2020.03.025

Figure Lengend Snippet: KEY RESOURCES TABLE

Article Snippet: 168 overlapping 15 residue peptides corresponding to MARV strain 200501379 Angola GP sequence were produced by JPT Peptide Technologies GmbH (Berlin, Germany).

Techniques: Recombinant, Infection, Staining, Expressing, Sequencing, Plasmid Preparation, Mutagenesis, Ligation, Peptide Microarray, Software, Flow Cytometry, High Throughput Screening Assay, Cytometry, Chromatography, Microscopy, Magnetic Beads

Journal: Blood

Article Title: Structural insights into inhibition of the bivalent menin-MLL interaction by small molecules in leukemia

doi: 10.1182/blood-2012-05-429274

Figure Lengend Snippet: Structure of the menin-MLL complex. (A) Sequence of the N-terminal fragment of MLL with MBM1 and MBM2 motifs. (B) Details of the menin-MBM1 interaction. Structure of the MBM1 is shown in stick representation (green carbon atoms) and MLL residues are labeled in blue. Menin is presented as a gray ribbon and selected side chains involved in contacts with MBM1 are shown as sticks (cyan carbon atoms); hydrogen bonds are shown as dashed lines. (C) The most significant contacts between MBM1 (green carbons) and selected menin side chains (cyan carbons). (D) Probing the MBM2-binding site on menin. MBM1 (shown in sticks) occupies a negatively charged central cavity on menin. The positions of D252 and L289 that were mutated to lysines are labeled. The electrostatic potential was calculated using APBS and mapped onto menin structure.41

Article Snippet: For crystallization experiments, 2.5 mg/mL of menin was incubated with MLL MBM1 peptide (MLL residues 4-15; GenScript) in a 1:1 molar ratio.

Techniques: Sequencing, Labeling, Binding Assay

Journal: Blood

Article Title: Structural insights into inhibition of the bivalent menin-MLL interaction by small molecules in leukemia

doi: 10.1182/blood-2012-05-429274

Figure Lengend Snippet: Crystal structure of the menin–MI-2 complex. (A) Details of MI-2 interaction with menin. Selected menin side chains are shown in sticks (cyan carbons) and hydrogen bonds are shown as dashed lines. (B) Menin-MI-2 complex determined at 1.56 Å resolution with corresponding 2Fo-Fc electron density map contoured at the 1σ level. (C) Diagram depicting VdW contacts and hydrogen bonds (dashed lines) between MI-2 and menin. (D) Superposition of MI-2 (green carbons) with a fragment of the MBM1 motif (MLL residues 9-13, gray carbons) in a menin-bound conformation.

Article Snippet: For crystallization experiments, 2.5 mg/mL of menin was incubated with MLL MBM1 peptide (MLL residues 4-15; GenScript) in a 1:1 molar ratio.

Techniques:

Journal: Blood

Article Title: Structural insights into inhibition of the bivalent menin-MLL interaction by small molecules in leukemia

doi: 10.1182/blood-2012-05-429274

Figure Lengend Snippet: Development of second-generation menin-MLL inhibitors. (A) Structures and activities of new compounds designed based on the structure of the menin–MI-2 complex. IC50 values for the inhibition of the menin-MBM1 complex are provided in parentheses. (B) FP experiments comparing activities of MI-2, MI-2-2 and MBM1 for disruption of the menin-MBM1 interaction demonstrating that MI-2-2 is a more potent inhibitor than an MLL-derived peptide. (C) Isothermal titration calorimetry showing the binding of MI-2-2 to menin. N represents a stoichiometry of binding. (D) Crystal structure of the menin–MI-2-2 complex determined at 1.27 Å resolution with corresponding 2Fo-Fc electron density map contoured at 1σ level. Water molecules were omitted for clarity. (E) Orthogonal dipolar interactions between MI-2-2 fluorine and backbone atoms of His181. The distances are shown in Å.

Article Snippet: For crystallization experiments, 2.5 mg/mL of menin was incubated with MLL MBM1 peptide (MLL residues 4-15; GenScript) in a 1:1 molar ratio.

Techniques: Inhibition, Disruption, Derivative Assay, Isothermal Titration Calorimetry, Binding Assay

Journal: Blood

Article Title: Structural insights into inhibition of the bivalent menin-MLL interaction by small molecules in leukemia

doi: 10.1182/blood-2012-05-429274

Figure Lengend Snippet: Small molecules targeting the MBM1 site efficiently disrupt bivalent menin-MLL interaction. (A) Fluorescence polarization experiments demonstrating displacement of MLL4-43 from menin by MI-2 and MI-2-2. Data represent mean values from 2 experiments ± SD. (B) Coimmunoprecipitation experiment in HEK293 cells transfected with Flag-MLL-AF9 showing that MI-2 and MI-2-2 disrupt the interaction of menin with MLL-AF9 in human cells. Input shows the levels of menin and MLL. The amount of menin bound to Flag-MLL-AF9 was detected by coimmunoprecipitation using anti-Flag Ab followed by immunoblotting using menin Ab. (C) Model of the disruption of bivalent MLL-menin interaction by MI-2-2 via binding to MBM1 site in menin.

Article Snippet: For crystallization experiments, 2.5 mg/mL of menin was incubated with MLL MBM1 peptide (MLL residues 4-15; GenScript) in a 1:1 molar ratio.

Techniques: Fluorescence, Transfection, Western Blot, Disruption, Binding Assay

Journal: Neuroscience

Article Title: α1b-Adrenergic Receptor Localization & Relationship to the D1-Dopamine Receptor in the Rat Nucleus Accumbens

doi: 10.1016/j.neuroscience.2017.11.053

Figure Lengend Snippet: Primary antibodies used for immunocytochemistry

Article Snippet: No bands were present when the cells were transfected with mock-DNA or DNA of another receptor (data not shown; Mitrano et al., 2010 ; 2014 ). table ft1 table-wrap mode="anchored" t5 caption a7 Antigen Immunogen Manufacturer Data Dilution Used α1bAR 15 amino acid peptide from the C-terminal residues of human α1bAR.

Techniques: Recombinant

Journal: Neuroscience

Article Title: α1b-Adrenergic Receptor Localization & Relationship to the D1-Dopamine Receptor in the Rat Nucleus Accumbens

doi: 10.1016/j.neuroscience.2017.11.053

Figure Lengend Snippet: Subcellular localization of D1Rs and α1bARs in the core and shell of the NAc. (A) Mean ± SEM of total D1R- or α1bAR-immunolabeled elements in the core and shell of the NAc using the single pre-embedding immunoperoxidase method. *p<0.01 when comparing D1R-containing dendrites to α1bAR-labeled dendrites in the core and shell. +p<0.001 when comparing D1R spines to α1bAR spines. #p<0.05 when comparing α1bAR-labeled unmyelinated axons to D1R axons. **p<0.001 when comparing α1bAR-labeled axon terminals to D1R terminals and ∘p<0.01 when comparing α1bAR-labeled glial elements to D1R glia. (B) Representative electron micrograph of α1bAR labeling in a dendrite, unmyelinated axon and an axon terminal in the NAc core. (C) Representative electron micrograph of α1bAR labeling in an unmyelinated axon and glial process in the NAc shell. (D) Representative electron micrograph of D1R labeling in 2 unmyelinated axons, 2 dendrites, one with a protruding spine synapsing on an unlabeled axon terminal. Den, dendrite; Sp, dendritic spine; UA, unmyelinated axon; AT, axon terminal; Ul., unlabeled. All scale bars = 0.5μm.

Article Snippet: No bands were present when the cells were transfected with mock-DNA or DNA of another receptor (data not shown; Mitrano et al., 2010 ; 2014 ). table ft1 table-wrap mode="anchored" t5 caption a7 Antigen Immunogen Manufacturer Data Dilution Used α1bAR 15 amino acid peptide from the C-terminal residues of human α1bAR.

Techniques: Immunolabeling, Labeling

Journal: Neuroscience

Article Title: α1b-Adrenergic Receptor Localization & Relationship to the D1-Dopamine Receptor in the Rat Nucleus Accumbens

doi: 10.1016/j.neuroscience.2017.11.053

Figure Lengend Snippet: α1bAR and D1R display a low degree of colocalization in the core and shell of the NAc. (A) The mean percent (± SEM) of α1bAR immunoperoxidase-containing elements that also contained immunogold labeling representing the D1R in both the core and shell of the NAc. *p<0.01 indicates in the NAc core, a significantly greater percentage of double-labeled dendrites vs. unmyelinated axons, axon terminals and glial elements. #p<0.05 indicates in the NAc shell, significantly more double-labeled dendrites than unmyelinated axons and axon terminals. (B) Representative electron micrograph from the NAc core with an immunoperoxidase-labeled dendrite representing α1bAR (arrowhead) and a D1R labeled dendrite (arrows point to PMB immunogold particles). (C) Electron micrograph of an immunoperoxidase-labeled α1bAR dendrite containing D1R PMB immunogold particles (arrows). (D) NAc shell, 4 α1bAR immunoperoxidase-labeled unmyelinated axons, α1bAR axon terminal (arrowhead), α1bAR dendrite with an unlabeled protruding spine. The single arrow points to a D1R-labeled unmyelinated axon immunogold particle. (E) Representative electron micrograph from the NAc core with α1bAR immunogold-labeled unmyelinated axons (single arrows point to PMB gold particles, double arrowhead indicates INT gold particles). (F) Electron micrograph from the NAc shell with 2 D1R immunoperoxidase-labeled dendrites, 2 α1bAR immunogold-labeled unmyelinated axons and one spine containing D1R immunoperoxidase labeling and immunogold labeling for the α1bAR (single arrows point to PMB gold particles, double arrowhead indicates INT gold particles). Den, dendrite; Sp, dendritic spine; UA, unmyelinated axon; AT, axon terminal; Ul., unlabeled. All scale bars = 0.5μm.

Article Snippet: No bands were present when the cells were transfected with mock-DNA or DNA of another receptor (data not shown; Mitrano et al., 2010 ; 2014 ). table ft1 table-wrap mode="anchored" t5 caption a7 Antigen Immunogen Manufacturer Data Dilution Used α1bAR 15 amino acid peptide from the C-terminal residues of human α1bAR.

Techniques: Labeling

Journal: Neuroscience

Article Title: α1b-Adrenergic Receptor Localization & Relationship to the D1-Dopamine Receptor in the Rat Nucleus Accumbens

doi: 10.1016/j.neuroscience.2017.11.053

Figure Lengend Snippet: Subsynaptic Distribution of α1bARs and D1Rs. (A) The mean percent (+/− SEM) of intracellular immunogold particles representing α1bARs in the NAc core and shell. *p<0.001 when comparing the mean percent intracellular immunogold particles to PMB particles on dendrites in both the core and shell; for axon terminals in the shell only. #p<0.01 when comparing the mean percent of intracellular to PMB gold particles in unmyelinated axons. Total number of elements examined containing only α1bAR immunogold: 69 dendrites (core), 52 (shell); 12 spines (core), 10 (shell); 189 unmyelinated axons (core), 141 (shell); 48 axon terminals (core), 50 (shell); 10 glial elements (core), 12 (shell). (B) The mean percent (+/− SEM) of intracellular immunogold particles representing D1Rs in the NAc core and shell. Total number of elements examined containing only D1R immunogold: 252 dendrites (core), 229 (shell); 76 spines (core), 56 (shell); 137 unmyelinated axons (core), 139 (shell); 21 axon terminals (core), 33 (shell); 3 glial elements (core), 4 (shell). (C) Mean percent (+/−SEM) of intracellular immunogold particles representing the α1bAR in single (Non-D1R) vs. double labeled elements (D1R) in the NAc shell; *p<0.05 when comparing intracellular immunogold particles for α1bAR in Non-D1R vs. D1R-containing elements. Total number of elements examined containing both α1bAR immunogold and D1R immunoperoxidase in the NAc shell: 77 dendrites; 12 spines; 21 unmyelinated axons; 17 axon terminals. (D) Mean percent (+/−SEM) of intracellular immunogold particles representing the D1R in single (Non-α1bAR) vs. double labeled elements (α1bAR) in the NAc shell; *p<0.01 when comparing intracellular immunogold particles for D1R in Non-α1bAR vs. α1bAR -containing elements. Total number of elements examined containing both D1R immunogold and α1bAR peroxidase in the NAc shell: 47 dendrites; 2 spines; 13 unmyelinated axons; 2 axon terminals.

Article Snippet: No bands were present when the cells were transfected with mock-DNA or DNA of another receptor (data not shown; Mitrano et al., 2010 ; 2014 ). table ft1 table-wrap mode="anchored" t5 caption a7 Antigen Immunogen Manufacturer Data Dilution Used α1bAR 15 amino acid peptide from the C-terminal residues of human α1bAR.

Techniques: Labeling