Journal: Biology Open

Article Title: Neuronal cell culture from transgenic zebrafish models of neurodegenerative disease

doi: 10.1242/bio.036475

Figure Lengend Snippet: Images of cultured 24 hpf Islet1:GFP zebrafish embryos stained with zebrafish-specific neuronal markers to confirm that the cell cultures contain various types of neurons. (A) An Islet1:GFP motor neuron within the cultures is positively stained (red) for the neuronal marker 39.4D5 (islet1 and islet2 homeobox). (B) Another Islet1:GFP motor neuron, and nearby islet1:GFP negative cells, are stained positively (red) for the neuronal cell surface marker Zn12, indicating the inclusion of other types of neurons in addition to motor neurons. Scale bar: 10 µm.

Article Snippet: Non-specific antibody binding was blocked by incubation by 5% goat serum (in PBS) prior to a 1 h incubation with one of the following primary antibodies: for islet 1 and islet 2 homeobox, 39.4D5 (1:50) and neuronal cell surface marker Zn12 (1:50), all obtained from the Developmental Studies Hybridoma Bank; for anti-polyglutamine (PolyQ) (Millipore, clone 5TF1-1C2|MAB1574).

Techniques: Cell Culture, Staining, Marker

Journal: PLoS ONE

Article Title: Differential Expression of Surface Markers in Mouse Bone Marrow Mesenchymal Stromal Cell Subpopulations with Distinct Lineage Commitment

doi: 10.1371/journal.pone.0051221

Figure Lengend Snippet: (A) An improved tet-inducible system for conditional expression of Large T-antigen. In the presence of Dex/Dox irtTA-GBD* fusion protein can be translocated to the nucleus and activate transcription of Large T. (B) MSCs were isolated from bone marrow of transgenic mice and expanded in the immortalization conditions (Phase Contrast, 20×). (C) Western blot showing induction of T-antigen in primary MSCs (“−“ cells before induction, “+DD” induced cells, “−DD” cells deinduced for 3 days). (D) Conditionally immortalized MSCs maintained differentiation potential in vitro into osteo-, adipo- and chondrogenic lineages. (E) Flow cytometry results for the markers highly expressed in MSCs, as shown by the staining with BD Lyoplate Antibody Screening Panel. Abbreviations: Dex – Dexamethasone, Dox – Doxycycline, irtTA – improved reverse tetracycline transactivator, GBD* - mutated glucocorticoid-binding domain, TAg – Large T-antigen, CAGGs and PGK – constitutively active promoters, tet-tk – tetracycline operator and minimal promoter, IRES – internal ribosome binding site, Puro and Hygro – puromycin and hygromycin resistance genes.

Article Snippet: BM MSCs were characterized using BD Lyoplate Mouse Cell Surface Marker Screening Panel (BD Biosciences, cat. 562208), which contains 176 monoclonal antibodies and correspondent isotype controls.

Techniques: Expressing, Isolation, Transgenic Assay, Western Blot, In Vitro, Flow Cytometry, Staining, Binding Assay