Sustained downregulation of CD62-L is mediated by PI3K and mTOR signaling.

<p>For FACS analysis, cells were treated and analyzed as described in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0068378#pone-0068378-g001" target="_blank">Figure 1</a>, while for qPCR analysis T-cells were recovered/enriched from co-cultures by MACS depletion. (A) Kinetics of CD62-L expression in the presence of the PI3K inhibitor, LY294002. (B) LY294002 has no effect on CD62-L loss in the first 6 hours after onset of T-cell activation but induces significant upregulation of CD62-L at later time points. (C) Kinetics of CD62-L expression in the presence of the mTOR inhibitor, Rapamycin. (D) Rapamycin has no effect on CD62-L loss in the first 6 hours after onset of T-cell activation but induces upregulation of CD62-L at later time points. (E) qPCR analysis of CD62-L mRNA expression level in TofB and TofDC at 24 and 72 hours. (F) qPCR analysis of KLF2 mRNA expression levels in TofB and TofDC at 24 and 72 hours. (G) qPCR analysis of CD62-L mRNA induction in both TofB and TofDC by Rapamycin and a detectable yet non-significant induction of KLF2 mRNA. (H) qPCR analysis of CD62-L and KLF2 mRNA induction in TofB and TofDC in the presence or absence of Ly294002. Data in B and D-H are means+SEM of 3–5 independent experiments.