Journal: Journal of Biochemistry

Article Title: Cell surface chondroitin sulphate proteoglycan 4 (CSPG4) binds to the basement membrane heparan sulphate proteoglycan, perlecan, and is involved in cell adhesion

doi: 10.1093/jb/mvy008

Figure Lengend Snippet: The CSPG4 and perlecan interaction is involved in cell adhesion and spreading. (A) MM200 cell adhesion on fibronectin coated surface was used as a positive control (i). MM200 cell adhesion on HCAEC perlecan coated surfaces, in the absence of antibody (ii), in the presence of antibodies against α2β1 integrin clone BHA2.1 (iii), against perlecan clone A74 (iv) and against CSPG4 clones B5 (v) and clone 9.2.27 (vi). Cells were stained for actin filaments with Rhodamine-phalloidin (i–vi) and imaged using confocal microscopy. White arrows point out the polymerized actin filaments. Scale bar represents 10 µm. (B) Live cell images using DIC were also taken after MM200 cell adhesion on HCAEC perlecan coated surfaces in the absence of antibody (i), in the presence of antibody against CSPG4 clones B5 (ii) and clone 9.2.27 (iii) for 1 h. Scale bar represents 50 µm. (C) Analysis of the perimeter of cells exposed to the perlecan coating for 1 h in the absence or presence of antibodies from (B). A minimum of 10 cells were analysed from three confocal images taken at random for each condition. Individual data points are indicated as well as mean ± standard deviation. (D) The number of cells adhered to the perlecan coating after 1 h in the absence or presence of anti-CSPG4 antibodies determined from the live cell DIC images in (C). Individual data points were indicated as well as mean ± standard deviation. ‘*’ denoted significant differences compared to cells plated on perlecan in the absence of antibodies (P < 0.05) analysed by one-way ANOVA. Data shown are representative of three independent experiments.

Article Snippet: Images were taken with differential interference contrast (DIC) microscopy (Nikon Eclipse TiE).

Techniques: Positive Control, Clone Assay, Staining, Confocal Microscopy, Standard Deviation

Journal: PLoS ONE

Article Title: Neurochemical Changes in the Mouse Hippocampus Underlying the Antidepressant Effect of Genetic Deletion of P2X7 Receptors

doi: 10.1371/journal.pone.0066547

Figure Lengend Snippet: A, B/Representative sections show rostral hippocampal DG areas in 1×1 sections of male wild type and P2rx7 knock out mice. Dark dots ( arrowheads ) represent the BrdU-positive cells (ImmPress-DAB-Ni staining). Camera lucida drawings of the same sections where the newly formed BrdU-labeled cells are indicated. The microscopic picture and drawing were taken at the same magnification (20X), and the bar indicates 100 µm. Histogram showing the average number of BrdU-positive cells in a rostral hippocampal DG area in the granule cell layer and in the 50-µm zone adjacent to its inner edge. C/We observed a significant difference (n = 5, p = 0.046) in the average number of labeled cells in the sections of P2rx7+/+ and P2rx7−/− mice.

Article Snippet: Confocal images were acquired at the same depth of the sections at same acquisition parameters with a Nikon A1R confocal system on an inverted Nikon Ti-E microscope (objective 20X DIC N1, numerical aperture 0.45 for large images and Plan Apo VC 60× Oil DIC N260X, numerical aperture 1.4 for details) equipped with NIS-Elements C software.

Techniques: Knock-Out, Staining, Labeling