Journal: Circulation Research

Article Title: Carcinoembryonic Antigen–Related Cell Adhesion Molecule 1 Inhibits MMP-9–Mediated Blood–Brain–Barrier Breakdown in a Mouse Model for Ischemic Stroke

doi: 10.1161/circresaha.113.301207

Figure Lengend Snippet: Figure 4. BBB breakdown is increased in Ceacam1-/- mice. Stability of the BBB during reperfusion (A, B, n=10 per group, representative microscopic images) and in sham mice (C, n=4 per group) was analyzed after intravenous injection of Evans blue. Semi-quantitative immunohistochemical analysis demonstrated higher BBB breakdown (A). This effect was independent of stroke size as shown in (B). No differences of Evans blue leakiness was observed in sham operated mice. Serum levels of Evans blue were measured as an internal control (D). (A) Representative microscopic images of brain sections after injection of Evans blue (red), scale bar: 2mm. The stroke area was identified with anti-MAP-2 antibodies (white; the stroke area did not stain positive for MAP2, black areas, marked with white dots). (B) Representative macroscopic images of Evans blue leakage (blue) combined with TTC-life/dead staining (the infarcted area is shown in white) of cortical (B, upper row) and striatal strokes

Article Snippet: ; Minneapolis, USA Ly6G / GR1 Rat polyclonal 1:50 AM26331PU-N Acris Antibodies, Herford Germany CD31 Rat Clone SZ31, IgG2a 1:50 DIA-310, Dianova, Hamburg, Germany MAP2 Rabbit polyclonal 1:100 sc-20172 Santa Cruz, Delaware, CA, USA NeuN Rabbit polyclonal 1:50 NBP1-77686 Acris Antibodies, Herford Germany hCEACAM1 Mouse Clone 4D1/C2, IgG1, 1:100 C. Wagener, UKE,Hamburg, Germany16 6 Primary antibody Donor species isotype dilution catalog number supplier hCEACAM1 Rabbit polyclonal 1:100 CfB-364RH Creative Biomart, Shirley, NY, USA hMMP-9 Rabbit polyclonal 1:100 3852S Cell Signaling, Danvers, MA, USA hCD68 Mouse Clone PGM1, IgG3 1:100 M0876, Dako, Glostrup, Denmark hMPO Rabbit polyclonal 1:100 AM09109PU-M Acris Antibodies, Herford Germany Antibodies used for Western blot analyses: Primary antibody Donor species isotype dilution catalog number supplier CEACAM1 Rabbit polyclonal 1:2000 Andrea Horst15 MMP-9 Goat polyclonal 0.1mg/ml AF909 R&D Systems,Inc.

Techniques: Injection, Immunohistochemical staining, Control, Staining

Journal: Experimental Animals

Article Title: Transmembrane protein modulates seizure in epilepsy: evidence from temporal lobe epilepsy patients and mouse models

doi: 10.1538/expanim.23-0092

Figure Lengend Snippet: The localization of transmembrane protein (TMEM230) in brain tissue by immunofluorescence assay. Among them, A–D reveals the localization of TMEM230 in the adult brain cortex, E–H reflect the localization of TMEM230 in the mouse brain cortex and I–L show the localization of TMEM230 in the mouse hippocampus. TMEM230 is labeled in green; microtubule-associated protein 2 (MAP2), a dendritic marker, is labeled in red; GFAP, an astrocyte marker, is labeled in blue. The cytoplasm of neurons is marked in yellow in D, H, and L. D, H, and L, represented by Merge, show the co-expression of TMEM230 (green stain), MAP2 (red stain) and GFAP (blue stain) in neurons. A–D showed that TMEM230 (green-stained) and MAP2 (red stain) were co-expressed in the cytoplasm of human cortical neurons (yellow stain), while TMEM230 (green stain) was not co-expressed with GFAP (blue stain) in the cytoplasm of human cortical neurons. E–H and I–L showed that TMEM230 (green-stained) co-expressed with MAP2 (red-stained) in the mouse cortex and the hippocampus. However, TMEM230 (green-stained) did not co-localize with GFAP (blue-blue-stained) in the cells.

Article Snippet: The following primary antibodies were used: TMEM230 antibody (rabbit polyclonal antibody, 1:100, Proteintech), a microtubule-associated protein 2 (MAP2) antibody (mouse polyclonal antibody, 1:100, Zhongshan Golden Bridge, Beijing, China), a glial fibrillary acidic protein (GFAP) antibody (chicken polyclonal antibody, 1:100, Zhongshan Golden Bridge), a VGLUT-1 antibody (mouse polyclonal antibody, 1:100, Proteintech) and a VGAT antibody (mouse polyclonal antibody, 1:100, Proteintech).

Techniques: Immunofluorescence, Labeling, Marker, Expressing, Staining

Journal: Journal of Dental Sciences

Article Title: Overexpression of sprouty 1 protein in human oral squamous cell carcinogenesis

doi: 10.1016/j.jds.2020.07.013

Figure Lengend Snippet: Western blot analyses: phosphor-ERK and total-ERK in human oral cancer cell lines as compared with a primary culture of normal oral mucosa (HOK). Upregulation of phosphor-ERK (normalized to total-ERK) in OECM1 and Ca922 cell lines as compared with HOK, and a slightly decreased expression in SAS as compared with HOK. Results were quantified using densitometric analysis, normalized to the level of β-actin, and expressed as a fold change relative to the normal oral mucosa. Bars represent means ± standard deviation of the mean (∗ P < 0.05). A representative result of three independent experiments is shown.

Article Snippet: Further, samples were analyzed using 10% SDS-PAGE (Sigma-Aldrich) gels, and the proteins were transmitted onto a PVDF membrane (Sigma-Aldrich) using Bio-Rad's transblot with primary antibodies against phosphor-ERK (Boster Biological Technology, CA, USA; Cat. No. P00104; 1:1000) and total-ERK (Boster Biological Technology; Cat. No. P00104; 1:1000), with species specificity for human tissues and an observed molecular weight of 42–44 kDa; and β-actin (Sigma-Aldrich; 1:1000), followed by horseradish peroxidase (HRP)-conjugated secondary antibodies (Sigma-Aldrich; 1:5000).

Techniques: Western Blot, Expressing, Standard Deviation

Journal: eLife

Article Title: Monoallelic CRMP1 gene variants cause neurodevelopmental disorder

doi: 10.7554/eLife.80793

Figure Lengend Snippet:

Article Snippet: Antibody , Anti-MAP2 (Rabbit polyclonal) , Covance , PRB−547C RRID: AB_2565455 , 1:5000.

Techniques: Variant Assay, Transfection, Construct, cDNA Library Assay, Expressing, Plasmid Preparation, Recombinant, Sequencing, Mutagenesis, Purification, Amplification, Software