Figure 3.

Binding of [³⁵S]GTPγS to ARFs 1, 5, and 6. (A) Samples (25 pmol) of ARF1 (●), ARF5 (▴), or ARF6 (■) were incubated for 20 min at 30°C with 4 μM [³⁵S]GTPγS and the indicated amount of ARF-GEP₁₀₀. (B) Samples (25 pmol) of ARF1, ARF5, or ARF6 were incubated with 4 μM [³⁵S]GTPγS for indicated time at 30°C without (○), or with 2.5 pmol of ARF-GEP₁₀₀ (■) or cytohesin-1 Sec7 (▴). Data are means ± SEM of values from triplicate assays. Findings were replicated at least three times with different protein preparations.