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. 2006 Nov 16;109(5):2190–2197. doi: 10.1182/blood-2005-01-031930

Figure 3.

Figure 3

Characterization of expression and function of full-length Helios. (A) Increased frequencies of DN thymocytes among cells that express full-length Helios. Data represent mice at 10 to 16 weeks after transplantation from 6 separate experiments. Asterisks indicate statistically significant differences between GFP control (n = 17) and Helios mice (n = 16) (DN1 and DN2, P < .001; DN3 and DN4, P < .05). DN1 thymocytes were analyzed as CD4−CD8−CD25−B220−Mac-1−CD44+ (B) Real-time reverse transcription-PCR analysis of endogenous Helios expression in thymocyte subsets sorted by FACS. Data shown are representative of 3 independent cDNA preparations (2 independent preparations for DN1) using mRNA from double-sorted cells. DN1 thymocytes were gated as CD4−CD8−CD25−B220−Mac-1−IL-7Rα −CD44+c-Kit+. cDNA from each subset was assayed in quadruplicate in each experiment. Error bars represent 1 SD from the mean. (C) CD4+ splenic T cells expressing either GFP or full-length Helios were stimulated in vitro with varying concentrations of anti-CD3 in the presence of irradiated APCs for 48 hours. Incorporation of tritiated thymidine into stimulated cells is shown. Error bars indicate ± SEM from quadruplicate wells from a representative experiment performed in triplicate.