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eji2476-sup-0001-FigureS1.ppt176K

Figure 1. Inducible disruption of the CD28 gene. (A) Targeting vector and treatment protocol. (B) Cell surface expression of CD28 on CD4+ T-cells 1 week after TM-treatment. Grey histograms show staining of CD28 on conventional CD28KO CD4 T-cells. (C) Absolute numbers (mean + SD) of conventional CD4 and CD8 T-cells are not affected after TM-induced gene deletion. A representative results of one out of three independent experiments are shown. (n = 5 mice per group per experiment)

Figure 2. Selective interruption of CD28 expression in mixed bone marrow chimeras (A) Irradiated RAG-/- deficient mice (10Gy as a single dose) were reconstituted with 2x107 T-cell-depleted mixed haematopoietic progenitors of BM (Bone Marrow) from iCD28KO and Thy1.1/CD28+/- donors at a 1:1 ratio. (B) Selective ablation of CD28 expression by TM-treatment (- 3 weeks) on iCD28KO-derived cells. One out of three independent experiments are shown.

Figure 3. Inducible deletion of CD28 reduces turnover of CD4 T-cells. (A)LN and spleen CD4 T-cells from iCD28KO mice were analyzed after 3 weeks of TM-treatment for expression of the proliferation marker Ki-67. (B) Ki-67+ conventional CD4 T-cells of reconstituted iCD28KO and Thy1.1/CD28+/- cell populations from RAG-/- recipients, 3 weeks after TM-induced CD28 deletion are compared to oil-administered recipients. Data from one out of two experiments are shown as mean+SD, (n = 5, per group and per experiment). *p < .05, **p < .01, ***p < .001, unpaired t test.

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