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. 2005 Jul;16(7):3211–3222. doi: 10.1091/mbc.E04-12-1065

Figure 8.

Figure 8.

GRASP65-depleted cells are arrested in mitosis before cell death. (A) HeLa cells transfected with scrambled control or GRASP65-specific siRNA were trypsinized at various time after transfection, stained with propidium iodide, and subjected to FACS analysis. The percentage of cells with 4N DNA content is shown. (B) The propidium iodide–stained cells prepared for the experiment described in A were analyzed for their sub-G1 (dead or damaged cells) content. The percentage of propidium iodide signal corresponding to dead or damaged cells is shown. (C) The mitotic index (positive staining with an antibody to phospho-histone H3) was determined by immunofluorescence 48 h after knockdown transfection from scrambled control-transfected or GRASP65-depleted cells. The mitotic index was also determined from knockdown and control-transfected cells that have been incubated for 16 h with 20 μM Z-VAD. (D) Mitotic GRASP65-depleted HeLa cells were left untreated or incubated with 20 μM Z-VAD for 16 h and stained with anti-α-tubulin antibody to reveal the organization of the mitotic spindle.