Figure 5.

Nocodazole-induced changes in Golgi organization are not perturbed by GRASP65 depletion. HeLa cells were transfected with the scrambled control or the GRASP65-specific siRNA. Forty-eight hours after transfection, cells were treated with 25 μg/ml nocodazole 90 min (Nocodazole) and either fixed or washed with medium to remove nocodazole and allowed to recover for 3 h (Nocodazole washout). The cells were processed for immunofluorescence microscopy using antibodies to GRASP65 to confirm the knockdown, to the cis-Golgi marker giantin and to the TGN marker TGN 46 to monitor the disassembly and reassembly of the cis- and trans-specific cisternae of the Golgi apparatus.