Imaging the intra-Golgi transport of VSVG under 15° and 20°C temperature block. HeLa cells transiently expressing the RUSH reporter, VSVG-SBP-GFP, were first incubated with nocodazole to induce the formation of ministacks. (A and B) Cells were subsequently treated with biotin and CHX in the presence of nocodazole at 15°C (A) or 20°C (B) for 90 or 120 min, respectively. Cells were fixed and immunostained for endogenous giantin before side-averaging. The organization of panels is similar to that of Fig. 4 C. Scale bar, 200 nm.