PMC:17824 / 8436-9486 JSONTXT

{"project":"2_test","denotations":[{"id":"11178126-10371511-4389688","span":{"begin":119,"end":121},"obj":"10371511"}],"text":"Hybridization in situ\nHybridization was conducted in situ to analyze cytokine mRNA expression, as previously detailed [10]. Briefly, synthetic oligonucleotide probes (Table 1) — TNF-α, IFN-γ, IL-1β, and IL-12 (the gift of Dr Tomas Olsson, Karolinska Institute, Stockholm, Sweden) — were labeled at the 3' end using terminal deoxynucleotidyl transferase (Advanced Biotechnologies, Leatherhead, UK) and [35S]ATP (Dupont Scandinavia, Stockholm, Sweden). Sections (6 μm thick) of freshly frozen synovial tissues were thaw-mounted onto slides and hybridized with 1 × 106 cpm of labeled probe per 100 μl hybridization mixture. After emulsion autoradiography, development, and fixation, the coded slides were examined by dark-field microscopy for positive cells, which were defined as those containing \u003e15 silver grains in a star-like distribution. The intracellular distribution of the grains was checked by light microscopy at higher magnification. The data were expressed as the number of cells (mean ± SEM) expressing mRNA per mm2 of the tissue section."}

{"project":"Colil","denotations":[{"id":"T22","span":{"begin":119,"end":121},"obj":"10371511"}],"namespaces":[{"prefix":"_base","uri":"http://pubannotation.org/docs/sourcedb/PubMed/sourceid/"}],"text":"Hybridization in situ\nHybridization was conducted in situ to analyze cytokine mRNA expression, as previously detailed [10]. Briefly, synthetic oligonucleotide probes (Table 1) — TNF-α, IFN-γ, IL-1β, and IL-12 (the gift of Dr Tomas Olsson, Karolinska Institute, Stockholm, Sweden) — were labeled at the 3' end using terminal deoxynucleotidyl transferase (Advanced Biotechnologies, Leatherhead, UK) and [35S]ATP (Dupont Scandinavia, Stockholm, Sweden). Sections (6 μm thick) of freshly frozen synovial tissues were thaw-mounted onto slides and hybridized with 1 × 106 cpm of labeled probe per 100 μl hybridization mixture. After emulsion autoradiography, development, and fixation, the coded slides were examined by dark-field microscopy for positive cells, which were defined as those containing \u003e15 silver grains in a star-like distribution. The intracellular distribution of the grains was checked by light microscopy at higher magnification. The data were expressed as the number of cells (mean ± SEM) expressing mRNA per mm2 of the tissue section."}