Journal: Experimental & Translational Stroke Medicine

Article Title: Neuronal precursor cell proliferation in the hippocampus after transient cerebral ischemia: a comparative study of two rat strains using stereological tools

doi: 10.1186/2040-7378-2-8

Figure Lengend Snippet: Infarct distribution in SDs and SHRs . Representative examples are shown of the infarct distribution after 90 minutes of tMCAo in the SD and SHR strains using NeuN and ED-1 immunohistochemistry. The ED-1 (brownish red rows) was visualized with NovaRed ® , whereas NeuN stain was developed with nickel-enhanced DAB (grey rows). A and B depict the infarct pattern we found in SDs. Usually, the infarcts were restricted to the subcortical structures in SDs encompassing both the MCA and AChA/HTA territories (4 SDs) ( A ) or the AChA/HTA territories alone (5 SDs) ( B ) Three of the tMCAo SDs had no histological signs of neuronal loss on the NeuN stain. C , D and E illustrate the infarct distribution in SHRs. In most of the SHRs subjected to ischemia the infarcted area included the neocortex (8 out of 11 SHRs). C shows an example of ischemia affecting the ACA, MCA and AChA/HTA (2 SHRs). The majority of animals had ischemia in the MCA and AChA/HTA territories (8 SHRs) ( D ). Only one SHR had a small ischemic lesion solely affecting the AChA/HTA territories ( E ). ACA , anterior cerebral artery; AChA , anterior choroidal artery; DAB , 3,3'-diaminobenzidine; HTA , hypothalamic artery; MCA , middle cerebral artery; NeuN , neuronal nuclei; SDs , Sprague-Dawley rats; SHRs , spontaneously hypertensive rats; tMCAo , transient middle cerebral artery occlusion.

Article Snippet: The sections were finally incubated with an avidin-biotin-peroxidase complex kit (Cat. no. PK-6100, Vector Laboratories, Burlingame, CA, USA) for 1 hour at room temperature before the development with nickel-enhanced DAB or NovaRed ® (Cat. no. SK-4100 and SK-4800, Vector Laboratories, Burlingame, CA, USA).

Techniques: Immunohistochemistry, Staining

Journal: eLife

Article Title: Predicted glycosyltransferases promote development and prevent spurious cell clumping in the choanoflagellate S. rosetta

doi: 10.7554/eLife.41482

Figure Lengend Snippet: FITC-labelled jacalin binds the apical and basal poles of wild type single cells ( B ) and becomes enriched in the ECM in the center of rosettes ( A, B’ boxed region from A ). Although FITC-jacalin staining appeared normal at the apical poles of Jumble ( C ) and Couscous ( D ) mutant cells, FITC-jacalin staining at the basal poles of cells was undetectable in cells grown either in the absence (-RIFs; C, D ) or presence (+RIFs; C’, D’ ) of RIFs. Arrows mark the apical pole and arrowheads mark the basal pole. ( E ) Cartoon depicts how jacalin fluorescence was measured. Starting with micrographs of FITC-jacalin stained cells, a line was drawn tracing from one edge of the collar around the cell body to the other edge of the collar, and the underlying fluorescent signal was normalized for cell size and background intensity. ( F ) The average normalized fluorescence intensity of jacalin measured in at least 59 cells for each condition was graphed against the normalized length of the cell body (n = 2 biological replicates). Jumble and Couscous -/+RIFs have reduced jacalin binding at the basal pole compared to wild type -/+RIFs. Gray shadows indicate 95% confidence intervals. Scale bars = 5 μm.

Article Snippet: Other , FITC-labelled jacalin , , Vector Labs , RRID: AB_2336460 ; Vector Labs: Cat. No.FLK-4100 , (1:400).

Techniques: Staining, Mutagenesis, Fluorescence, Binding Assay

Journal: eLife

Article Title: Predicted glycosyltransferases promote development and prevent spurious cell clumping in the choanoflagellate S. rosetta

doi: 10.7554/eLife.41482

Figure Lengend Snippet: ( A–F ) Biotinylated-jacalin labelled with streptavidin Alexa Fluor 647 has the same localization pattern in the absence ( A, C, E ) and in the presence ( B, D, F ) of RIFs as that observed with FITC-labelled jacalin . In wild type cells, jacalin binds the apical and basal poles of single cells ( A ) and becomes enriched at the center of wild type rosettes ( B ). In the mutants Jumble ( C, D ) and Couscous ( E, F ), jacalin staining was severely reduced at the basal poles both in the absence ( C, E ) and in the presence ( D, F ) of RIFs, while the apical pole staining appeared similar to wild type single cells. ( G, I ) Transfection of Jumble ( G ) and Couscous ( I ) with mTFP alone did not restore jacalin localization to the basal pole. Shown here in the presence of RIFs. ( H, J ) However, Jumble ( H ) and Couscous ( J ) complemented with mTFP-jumble or couscous-mTFP , respectively, form rosettes with jacalin localized in the center as observed in wild type rosettes. Arrows mark the apical pole and arrowheads mark the basal pole. Scale bar = 5 μm.

Article Snippet: Other , FITC-labelled jacalin , , Vector Labs , RRID: AB_2336460 ; Vector Labs: Cat. No.FLK-4100 , (1:400).

Techniques: Staining, Transfection