Figure 3.

Recruitment of AP-1 onto the TGN with ARF·GTP. Digitonin-permeabilized normal rat kidney cells were mixed with 5 mg/ml gel-filtered rat liver cytosol and either 100 μM GTPγS (a and b) or 1 mM GTP (c and d) or 1 mM GTP and 4 μM ARF1 (e–i) and then incubated at 37°C for 20 min. After washing and fixation, the cells were analyzed with the anti-α-mannosidase II mAb 53FC3 (a, c, and e), affinity-purified anti-adaptor β subunit antibody GD/1 (b, d, f, and h), the anti-TGN38 mAb 2F7.1 (g), or affinity-purified anti-AP-1 γ subunit antibody AE/1 (i).