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. Author manuscript; available in PMC: 2013 Dec 15.
Published in final edited form as: J Immunol. 2012 Nov 5;189(12):5886–5895. doi: 10.4049/jimmunol.1103092

Figure 2. Naïve human T cell migration to S1P requires intact AKT signaling.

Figure 2

(A) Cells pretreated with ManLAM do not modulate surface CCR7 or S1P1 expression as determined by flow cytometry (grey filled histogram = isotype control, black = PBS treated, grey = ManLAM treated; representative image). (B) Quantification of surface receptor expression as in (A) (n=3 donors assayed in duplicate, mean± s.e.m.). (C) Western blotting analysis of human T cells pretreated with ManLAM or PBS for 2h, then stimulated with S1P or vehicle for 5 minutes. Percent phosphorylated AKT was determined by creating a ratio of densitometry of phospho-AKT Ser473 to total AKT. (Representative blot, n=2 separate experiments with 3 donors). (D) Pretreatment with 10μM Ly294002 for 1h inhibits human T cell migration to S1P. (*P=0.05 Student’s t test, n=2 donors assayed in duplicate, mean ± s.e.m.).