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. 1999 Jan 19;96(2):493–498. doi: 10.1073/pnas.96.2.493

Figure 2.

Figure 2

Effect of pretreatment of porcine follicular membranes with neutralizing arrestin antibodies on desensitization of hCG-stimulated AC activity. (A) Membranes (≈30 μg) were subjected to a preincubation (15 min at room temperature; 1 h at 4°C) with indicated final concentrations (in 50 μl reaction volume) of arrestin antibody A9C6 or NIS, followed by the two-stage desensitization reaction at 30°C, which consisted of a 40-min stage 1 desensitization incubation followed by a 5-min AC assay. Figure shows basal (BSA/BSA), full hCG-stimulated (BSA/hCG), and hormone-desensitized hCG-sensitive AC activity (hCG/hCG). Results are means ± SEM of quadruplicate determinations of a single assay and are representative of four separate experiments. Percentage of desensitization (% D) represents the reduction in hCG-stimulated AC activity when incubations contained hCG in stage 1 compared with activity measured with BSA in stage 1 over basal (BSA/BSA) AC activities. (B) Composite effect of different concentrations of arrestin antibody A9C6 or NIS at indicated final concentrations on percentage of desensitization of hCG-stimulated AC activity. Results are means ± SEM of four separate experiments. Remaining details are as in A. ∗, Differences between percentage desensitization values with arrestin antibody and NIS are significantly differently (P < 0.05). (C) Membranes were preincubated with indicated final concentrations of NIS, A9C6, F4C1, or both A9C6 and F4C1, as in A. Results are means ± SEM of quadruplicate determinations in a single assay.