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Real-time quantitative PCR analysis of mouse immature CD4SP thymocytes

ABSTRACT: RNA was isolated with the RNeasy kit (Qiagen) from electronically sorted immature CD4SP thymocytes. RNA was reverse transcribed into cDNA by oligo(dT) priming with the QuantiTect Reverse transcription kit (Qiagen). The expression of 84 key genes involved in apoptosis, or programmed cell death was performed with mouse apoptosis RT2ProfilerTM PCR array system (PAMM-012 E; SABiosciences). PCR Array data were analyzed by the Ct data analysis method. qPCR gene expression profiling. CD4SP thymocytes from WT and Ik7Tg mice were used.

ORGANISM(S): Mus musculus

SUBMITTER: Jung-Hyun Park

PROVIDER: E-GEOD-49270 | biostudies-arrayexpress |

REPOSITORIES: biostudies-arrayexpress

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Ikaros is required to survive positive selection and to maintain clonal diversity during T-cell development in the thymus.

Tinsley Kevin W KW Hong Changwan C Luckey Megan A MA Park Joo-Young JY Kim Grace Y GY Yoon Hee-Won HW Keller Hilary R HR Sacks Andrew J AJ Feigenbaum Lionel L Park Jung-Hyun JH

Blood 20130801 14

The zinc-finger protein Ikaros is a key player in T-cell development and a potent tumor suppressor in thymocytes. To understand the molecular basis of its function, we disabled Ikaros activity in vivo using a dominant negative Ikaros transgene (DN-IkTg). In DN-IkTg mice, T-cell development was severely suppressed, and positively selected thymocytes clonally expanded, resulting in a small thymus with a heavily skewed T-cell receptor (TCR) repertoire. Notably, DN-IkTg induced vigorous proliferatio ...[more]

PMID: 23908463

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