The RNA-Binding Protein KSRP Modulates Cytokine Expression of CD4+ T Cells
Inactivation of the KSRP gene enhances proliferation of polyclonally stimulated CD4+ T cells. Magnetic bead separation was used to isolate untouched CD4+ and CD8+ T cells from KSRP+/+ (WT) and KSRP-/- (KO) mice. T cells were polyclonally stimulated with plate-bound anti-CD3- (1 μg/ml) and anti-CD28- (2 μg/ml) specific antibodies for 96 h. In order to assess the proliferation of CD4+ (a) and CD8+ (b) T cells, 3H thymidine (0.5 μCi/well) was applied for the last 16-18 h of culture. 3H thymidine uptake is presented. Data are the ( animals per genotype) and are pooled from three independent experiments ( and ; ns = not significant, two-way Anova/Bonferroni test). Expression of the indicated surface markers in polyclonally stimulated CD4+ and CD8+ T cells isolated from KSRP+/+ (WT) and KSRP-/- (KO) mice was assessed by flow cytometry. One representative FACS analysis from two to three independent experiments is shown demonstrating CD4 (c) (APC-anti-CD4), CD8 (d) (eFluor450-anti-CD8), and CD25 (FITC-anti-CD25 mAb) expression. Data are the ( animals per genotype) and are pooled from 2 to 3 independent experiments (; ns = not significant, unpaired -test).
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