Journal: Cells

Article Title: Regulation of the Intestinal Extra-Adrenal Steroidogenic Pathway Component LRH-1 by Glucocorticoids in Ulcerative Colitis

doi: 10.3390/cells11121905

Figure Lengend Snippet: Dexamethasone induces cortisol and LRH-1, enhancing GR-binding to the NR5A2 gene promoter in colonocytes. ( A ) The CCD841CoN colonocyte cell line was stimulated with dexamethasone (100 nM) for 3, 6 and 24 h, and cortisol levels were determined in supernatants by ELISA; ( B ) Cells upon Dex and/or GR antagonist RU-486 (10 µM) treatment for 8 h were used to evaluate mature ( left ) and precursor ( right ) NR5A2 mRNA by RT-qPCR, and protein content by immunoblot; ( C ) Band quantification was performed by densitometry analysis and normalized to β-actin protein (% of control); ( D ) Cells stimulated with Dex for 2 h were analyzed for binding of GR to the NR5A2 promoter by ChIP-qPCR; For each immunoprecipitated sample, the statistical analyses were performed with respect to an unrelated region from the GAPDH gene promoter. Results are expressed as % input ± SEM. One-way ANOVA with Tukey post-test was performed. ** p < 0.01, *** p < 0.001, **** p < 0.0001. C: control, Dex: dexamethasone, RU: RU-486, GRE: glucocorticoid responsive element, IgG: immunoglobulin G, nt: nucleotides; n = 4.

Article Snippet: Cortisol concentrations were measured in supernatants of human colonic biopsies cultured in 350 ul with different treatments (Dex, RU-486, SR-1848) using the ELISA kit DetectX Cortisol Enzyme Immunoassay Kit (Arbor Assays, Ann Arbor, MI, USA, Cat. K003-H5), as detailed in the .

Techniques: Binding Assay, Enzyme-linked Immunosorbent Assay, Quantitative RT-PCR, Western Blot, Control, ChIP-qPCR, Immunoprecipitation

Journal: Cell Reports Medicine

Article Title: A GPC2 antibody-drug conjugate is efficacious against neuroblastoma and small-cell lung cancer via binding a conformational epitope

doi: 10.1016/j.xcrm.2021.100344

Figure Lengend Snippet:

Article Snippet: Antibodies utilized were normal rabbit IgG (4 μL; Cell Signaling, #2729), Histone H3 (D2B12) XP Rabbit mAb (10 μL; Cell Signaling, #4620), and N-MYC (4 μL; Active Motif, #61185).

Techniques: Control, Western Blot, Virus, Cell Culture, Recombinant, Expressing, Transfection, Cell Viability Assay, Caspase-Glo Assay, Luciferase, Mutagenesis, Antibody Labeling, Sonication, Chromatin Immunoprecipitation, SYBR Green Assay, Fluorescence, Quantitation Assay, Conjugation Assay, Gene Expression, Plasmid Preparation, shRNA, Software