Fig. 6
ROR1P(841)A has impaired capacity to associate with cortactin, induce cortactin phosphorylation, or enhance chemokine-directed MEC1-cell F-actin polymerization. a Schematic depicts the structure of ROR1 protein with different domains. b ΔPRD represents the truncated form of ROR1 without its PRD. c Amino-acid sequences of the PRD of ROR1. Asterisks indicate the proline (P) amino-acid residues that had been substituted with alanine (A). d Interaction of ROR1 with cortactin was determined by immunoblot analysis of anti-ROR1 ip from lysates of MEC1 (Ctrl), MEC1-ΔPRD, or MEC1-ROR1 (W/T) cells as indicated on the top. e Interaction of ROR1 with cortactin was confirmed by immunoblot analysis of anti-ROR1 ip from lysates of MEC1, MEC1-ΔPRD, MEC1-ROR1 (W/T), or MEC1 cells transfected with each of the various mutated forms of ROR1, as indicated on the top (upper panel). In the lower panel is an immunoblot of the whole-cell lysates of the MEC1 (Ctrl), MEC1-ΔPRD, MEC1-ROR1 (W/T), or MEC1 cells transfected with each of the various mutated forms of ROR1, as indicated on the top, and probed with anti-cortactin or anti-pCortactin (Y421) antibody. Expression and tyrosine phosphorylation of cortactin (Y421) was determined in the whole-cell lysates. f MEC1 (Ctrl), MEC1-ROR1 (W/T), or MEC1 cells transfected with each of the various mutated forms of ROR1, as indicated at the bottom, were examined for F-actin polymerization in the absence (–) or presence (+) of chemokine CCL21 (100 ng/ml). Data are shown as mean ± S.D. from three independent experiments, (n = 3). P < 0.05; P < 0.01, as calculated using one-way ANOVA with post-hoc Tukey HSD test
