Periciliary delivery is actin dependent and old gp135 is cleared from the periciliary region in a microtubule-dependent manner. (A) Newly delivered gp135 (BG488, green) after 0–20 min at 37°C in untreated or latrunculin A (latA)-treated MDCK-S cells (ZO1, blue). (B) Mean line scan intensity of newly delivered gp135, as in Fig. 2 B, demonstrates inhibition of periciliary delivery in latA-treated cells. All values were normalized to highest mean value of BG488 intensity at 10 min in untreated cells. (C) Old surface gp135 (BG549) after 0–10 min at 37°C in untreated or nocodazole-treated MDCK-S cells. (D) Mean line scan intensity of old gp135, as in Fig. 2 B, demonstrates inhibition of gp135 clearing from the periciliary region in nocodazole-treated cells. (B and D) 0 min, dashed line; 10 min, solid line. n = 15 cells per condition. For both experiments, data are representative of three independent experiments. Bars, 10 µm.