Figure 8.

CXCR3 antagonists inhibit CXCR3-mediated phosphorylation of p44/p42 extracellular signal-regulated kinase (ERK) and protein kinase B (PKB)/Akt in human T cells. Aliquots of activated T cells (1 × 10⁶ cells/500 μl) were left untreated or stimulated at 37° with 1 nm CXCL11 or CXCL12 for 2 min in the absence or presence of 30 min pre-treatment with T487 or NB-74330 at 1 μm and 100 nm, respectively. Cell lysates were resolved by SDS–PAGE, transferred to nitrocellulose membranes, and immunoblotted with a phospho-specific Erk or Akt antibody with affinity for the active Ser473-phosphorylated form of Akt and proteins were visualized with enhanced chemiluminescence (a). The blots were stripped and reprobed with anti-Erk-1 antibody to verify equal loading and efficiency of protein transfer (lower panel). Phosphorylation of Akt and p44/p42 ERK was quantified by densitometry and corrected for total Erk 1 expression on stripped blots (b). The data are derived from a single experiment representative of at least three others using cells from different donors.