PMC:3291650 / 35375-36071 JSONTXT

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{"project":"testone","denotations":[{"id":"T16088","span":{"begin":576,"end":586},"obj":"Gene_expression"},{"id":"T16087","span":{"begin":96,"end":106},"obj":"Gene_expression"},{"id":"T16086","span":{"begin":571,"end":574},"obj":"Protein"},{"id":"T16085","span":{"begin":562,"end":565},"obj":"Protein"},{"id":"T16084","span":{"begin":501,"end":504},"obj":"Protein"},{"id":"T16083","span":{"begin":107,"end":110},"obj":"Protein"},{"id":"T16082","span":{"begin":82,"end":85},"obj":"Protein"},{"id":"T16081","span":{"begin":70,"end":80},"obj":"Protein"},{"id":"T16080","span":{"begin":57,"end":60},"obj":"Protein"},{"id":"T16079","span":{"begin":22,"end":25},"obj":"Protein"}],"relations":[{"id":"R12117","pred":"themeOf","subj":"T16081","obj":"T16087"},{"id":"R12118","pred":"equivalentTo","subj":"T16082","obj":"T16081"},{"id":"R12119","pred":"themeOf","subj":"T16085","obj":"T16088"},{"id":"R12120","pred":"themeOf","subj":"T16086","obj":"T16088"}],"text":"In vivo intracellular GrB and IFNγ staining of activated CD8+ T cells\nGranzyme B (GrB) and IFNγ expressing CD8+ T cells were determined by treating the mice intranasally with 50 µg Brefeldin A (Sigma) as previously described [86]. 6 h later, BAL and lungs were isolated and single cell suspensions were prepared from the lung in the presence of 3 µg/ml Brefeldin A. Cells were stained, fixed and permeabilized (Cytofix/Cytoperm, BD Biosciences) according to the manufacturer's instructions. Activated CD8+ T cells were analyzed by flow cytometry based on CD62lo CD3+ and CD8+ expression. Live/Dead fixable aqua dead cell stain kit (Molecular Probes) was used to discriminate live from dead cells."}