Fig. 7.

Expression of Myc-RhoB Q63L does not impair CXCR2 recycling. ¹²⁵I-CXCL8 binding in HEK293 cells stably expressing CXCR2 and transfected with empty vector or Myc-RhoB Q63L. Cells were pretreated with 20 μg/ml cycloheximide for 30 minutes and placed in the presence of 20 μg/ml cycloheximide throughout the experiment. Cells were stimulated with vehicle (total binding), 100 ng/ml CXCL8 for 30 minutes, CXCL8 was removed, and cells were allowed to recover for 0 minutes, 30 minutes or 60 minutes. Cells were incubated with 0.1 nM ¹²⁵I-CXCL8 (specific activity =2200 Ci/mmol) for 2 hours, washed to remove non-specific binding, and subjected to γ-counting as described in Materials and Methods. Values represent three independent experiments and are shown as percent of total binding ± s.e.m. Data shown are representative from three separate experiments.