Journal: Cell

Article Title: Structural and Functional Analysis of the D614G SARS-CoV-2 Spike Protein Variant

doi: 10.1016/j.cell.2020.09.032

Figure Lengend Snippet:

Article Snippet: All specific SPR binding sensorgrams were double-reference subtracted as reported previously ( ) and the kinetic parameters were obtained by globally fitting the double-reference subtracted data to a 1:1 binding model with mass transport limitation using Biacore T200 Evaluation software v 3.1 (GE Healthcare).

Techniques: Binding Assay, Recombinant, Transfection, Luciferase, Bradford Protein Assay, Plasmid Preparation, Software, Single Particle, Microscopy

Journal: Proceedings of the National Academy of Sciences of the United States of America

Article Title: Identification of a tumor-promoter cholesterol metabolite in human breast cancers acting through the glucocorticoid receptor

doi: 10.1073/pnas.1707965114

Figure Lengend Snippet: Binding of the indicated ligands to ERα and LXRs, extinction of GR and LXRβ expression in the indicated BC cells and evidence that OCDO does not stimulate cell proliferation through the LXRβ. (A–F) Representative SPR sensorgrams from three experiments showing the binding of a series of concentrations of OCDO, 22(R)HC, or 17β-estradiol to the indicated receptor-LBD, captured on a Biacore sensor chip. Concentrations: 6.25 µM (red), 12.5 µM (green), 25 µM (dark blue), 50 µM (pink), 100 µM (light blue). Each sensorgram (expressed in RUs as a function of time in seconds) represents a differential response where the response on an empty reference channel (Fc1) was subtracted. The affinity constant (KD) is determined at equilibrium by the BIAevaluation software. (G and H) GR expression (G) or LXRβ expression (H) in MCF7 or MDA-MB-231 cells was knocked down using either shRNA against the GR (clones shGR5 and shGR6), or the LXRβ (clones shLXR3 and LXR4), or control shRNA (shC) and the expression of receptors was analyzed by immunoblotting. The blots are representative of three experiments. (I) Effect of solvent vehicle (control) or OCDO (5 or 10 µM) on the proliferation of the indicated tumor cells using a colorimetric BrDU immunoassay. (J) Effects of solvent vehicle (control) or OCDO (5 µM) or DEX 0.1 µM, or OCDO (5 µM) + DEX 0.1 µM on the proliferation of the indicated tumor cells using a colorimetric BrDU immunoassay. (I and J) Data are the means (±SEM) of five separate experiments, (n = 8), **P < 0.01, ***P < 0.001, one-way ANOVA, Tukey’s posttest. ns, not significant.

Article Snippet: Binding parameters were obtained by fitting the overlaid sensorgrams with the 1:1 Langmuir binding model of the Biacore T200 Evaluation software v2.0 or the Steady State affinity model of the BIAevaluation software v3.1.

Techniques: Binding Assay, Expressing, Software, shRNA, Clone Assay, Control, Western Blot, Solvent