Figure 1.

Depletion of GRASP65 by siRNA promotes cell death. (A) The sequences of the human GRASP65 gene used for designing SiRNA probes are shown (exact location is indicated). GR65–1–scrambled represents a scrambled version of GR65–1 used as control. (B) HeLa cells were transfected with GR65–1, GR65–2, GR65–1–scrambled or buffer. After 48 h, the level of GRASP65 was analyzed by immunoprecipitation/western blotting from total lysate, whereas the levels of the Golgi proteins GM130, Golgin 97, and GRASP55 were detected by Western blot analysis of the same lysates. (C) HeLa cells grown in six-well dishes were transfected with GR65–1, GR65–2, or GR65–1–scrambled. At the indicated time points, cells from two independent wells/per transfection were trypsinized and stained with trypan-blue to identify dead cells, followed by counting the live (i.e., trypan-blue negative) cells.