Figure 4.

Translocation of IRAP to the cell surface after treatment of the cells with insulin. (a and b) Effect of incubation time on the response to 10⁻⁷ M insulin. CHO-K1 (a) and recombinant HEK293 (b) cells were incubated with insulin for different time periods. (c and d) Effect of the insulin concentration on the binding of [³H]Ang IV. CHO-K1 (c) and recombinant HEK293 (d) cells were incubated with different concentrations of insulin for 6 min. Subsequently, all cells were further treated as mentioned in Table 1 (steps 1–3). Data refer to specific binding, expressed as a percentage of the control (no insulin). Values are the average±s.e.mean of three experiments with determinations performed in triplicate. Ang, angiotensin; CHO-K1, Chinese hamster ovary cells; HEK293, human embryo kidney cells; IRAP, insulin-regulated aminopeptidase.