Fig. 1.

Abl family kinases are required for chemokine-induced T cell migration and are activated by chemokines. (A) Primary wild-type (WT) and Abl/Arg null T cells were tested for their ability to migrate to either SDF-1α or CCL21 in transwell chambers. Data are means ± SEM (n = 5 experiments). (B) WT, Abl-deficient, Arg-deficient, and Abl and Arg doubly deficient (Abl/Arg null) T cells were tested for migration to CCL21. Data are means ± SEM (WT, n=4; Abl^−/−, n=3; Arg^−/−, n=4; Abl/Arg^−/−, n=3 experiments). (C) Mouse primary T cells were stimulated with SDF-1α or CCL21 for 30 s, and activation of Abl family kinases was detected by Western blotting analysis of the phosphorylation of CrkL (pCrkL, Y207). Blots were stripped and then analyzed for total CrkL protein. (D) WT or Abl/Arg null T cells were stimulated with SDF-1α and analyzed by Western blotting for the presence of pCrkL as described for (C). Total cell lysates were analyzed by Western blotting for Abl and Arg. Quantification of the amount of pCrkL normalized to that of total CrkL is shown in the bar graph below (n = 4 experiments). (E) Human H9 T cells that were untreated or were pretreated with STI571 were plated onto transwell inserts precoated with fibronectin. Migration of the cells to increasing amounts of SDF-1α is shown. Data are means ± SD of triplicate values. (F) H9 T cells that were untreated or were pretreated with STI571 were stimulated with SDF-1α at the indicated concentrations for 30 s, and lysates were analyzed by Western blotting for pCrkL as described for (C). Blots were then analyzed for total CrkL protein. Data are representative of three (C and E) or two (F) independent experiments. *P < 0.05, **P < 0.01, and ***P < 0.001.