Journal: Cell Reports

Article Title: The N-Terminal Domain of cGAS Determines Preferential Association with Centromeric DNA and Innate Immune Activation in the Nucleus

doi: 10.1016/j.celrep.2019.01.105

Figure Lengend Snippet:

Article Snippet: TransIT®-293 Transfection Reagent , Euromedex , Cat# MIR2706.

Techniques: Virus, Recombinant, Transfection, Protease Inhibitor, Lysis, Plasmid Preparation, SYBR Green Assay, Binding Assay, Control, Enzyme-linked Immunosorbent Assay, Software, FCAP Assay

Journal: International Journal of Molecular Sciences

Article Title: P2X4 Receptors Mediate Ca 2+ Release from Lysosomes in Response to Stimulation of P2X7 and H 1 Histamine Receptors

doi: 10.3390/ijms221910492

Figure Lengend Snippet: Colocalization of P2X4 receptors with endolysosomal markers. rP2X4-EGFP receptors were expressed in NRK cells, and live cells were imaged by confocal fluorescence microscopy 48 h post transfection after co-labeling lysosomes with a variety of different markers. These include ( A ) LAMP1-mCherry (LAMP1-mCh), which was co-transfected with rP2X4-EGFP, and ( B ) Texas Red-labeled Dextran (DexTR) incubated for 5 h and chased for 2 h at 37 °C. ( C ) Calculation of Pearson’s coefficient showed the high degree of colocalization between rP2X4 and LAMP1-mCh/DexTR. Results are the mean ± SEM from three independent experiments, total number of cells, n = 40. ( D ) Cells were incubated with Lysotracker (50 nM) for 10 min at 37 °C and, in ( E , F ), with a fluorescent substrate of cathepsin B, Magic Red TM (MR) for 5 min. Cells in ( F ) were imaged using TIRF microscopy to show only those rP2X4- and MR-positive compartments located within ~100 nm of the plasma membrane. Arrowheads indicate examples of compartments that are labeled only by rP2X4 or MR. Scale bars are 10 µm (5 µm for enlargements).

Article Snippet: TransIT-LT transfection reagent was from Geneflow (Lichfield, UK), ionomycin from Apollo Scientific (Stockport, UK), polyethylenimine (PEI) from Polysciences (Warrington, PA, USA), PBS tablets from Gibco (Dublin, Ireland), Magic Red TM kit from BioRad (Hercules, CA, USA), and nigericin (sodium salt) and monensin (sodium salt) from the Cayman Chemical Company (Ann Arbor, MI, USA).

Techniques: Fluorescence, Microscopy, Transfection, Labeling, Incubation, Clinical Proteomics, Membrane