Figure 1.

GM130 regulates the GA association of GRASP65, not its stability. (A) RNAi targeting sequences (GM130-1 and GM130-2) and their positions within the human GM130 cDNA are shown. (B) HeLa cells, transfected with the control or knockdown siRNA (GM130-1 siRNA) or short hairpin-expressing plasmids (GM130-1 shRNA), were harvested at the indicated time points after transfection. Western blot analysis was used to reveal the levels of GM130, GRASP65, and α-tubulin as a loading control. (C) Lysates of control and GM130-depleted cells were examined by Western blot analysis with antibodies to GM130, Golgin 97, GRASP55, and α-tubulin as a loading control. (D) Control or GM130-depleted cells were analyzed by immunofluorescence with antibodies to GRASP65, GRASP55, and Giantin 72 h after knockdown transfections. Scale bar, 10 μm.