Figure 1.

PLA₂ antagonists inhibit the nocodazole-induced formation of Golgi ministacks. Clone 9 rat hepatocytes were left untreated (A and B) or incubated at 4°C for 20 min before shifting to 37°C in 6 μg/ml nocodazole (Noc) for 2 h (C and D). To examine the effect of PLA₂ antagonists on ministack formation, we incubated the cells at 4°C for 20 min with 5 μM BEL (E and F), 25 μM AACOCF₃ (G and H), or 25 μM ONO-RS-082 (ONO; I and J) and then shifted the cells to 37°C in the continued presence of the same PLA₂ antagonist plus 6 μg/ml nocodazole for 2 h (E–H) or 1 h (I and J). All conditions included 2 μg/ml cycloheximide. Cells were fixed and processed for immunofluorescence microscopy (see MATERIALS AND METHODS) using a polyclonal antibody against the medial Golgi enzyme ManII (A, C, E, G, and I) and a monoclonal antibody against α-tubulin (B, D, F, H, and J).