Journal: Journal of Virology
Article Title: Genome Structure and Thymic Expression of an Endogenous Retrovirus in Zebrafish
Figure Lengend Snippet: Expression of ZFERV-related RNA in thymus. (A) Larval fish at 5 dpf were fixed and subjected to whole mount in situ hybridization with a DIG-labeled sense or antisense probe for the env gene, followed by colorimetric detection. (B and C) Contiguous sections (20 μm thick) of 3-month-old fish (right two panels) were subjected to in situ hybridization with probes as described in panel A. The section planes and main organs are indicated. Abbreviations: B, brain; E, esophagus; G, gill; H, heart; I, intestine; L, liver; M, muscle; T, thymus. The bilateral thymi hybridizing to the antisense probe are highlighted with arrows. (D) Sections (2 μm thick) of the thymus from 3-month-old fish were hybridized to fluorescein-labeled sense (left panel) or antisense (right panel) env probe, followed by fluorescence detection with the alkaline phosphatase-Fast Red system.
Article Snippet: For hybridization, samples were incubated with prehybridization buffer (4× SSC [600 mM NaCl, 60 mM sodium citrate; pH 7], 50% [vol/vol] formamide) at 37°C for 10 min, followed by incubation at 42°C for 16 h in hybridization buffer (50% [vol/vol] formamide, 4× SSC, 1× Denhardt solution [200 mg/liter each for Ficoll 400, polyvinylpyrrolidone, and bovine serum albumin], 100 μg of heparin/ml, 0.1% Tween 20, 1 mg of yeast Torula RNA [Sigma]/ml) containing fluorescein- or digoxigenin [DIG]-labeled RNA probe (0.2 μg/ml).
Techniques: Expressing, Fluorescence In Situ Hybridization, In Situ Hybridization, Labeling, Fluorescence