Survivin expression in factor-dependent MO7e cells.

Log phase MO7e cells growing in 20% FBS and 20 ng/mL GM-CSF (panel, lane 1) were washed and factor-starved for 24 hours in the absence of GM-CSF and FBS. Starved cells (panel, lane 2) were suspended in RPMI plus 10% FBS and cultured in the absence (panel, lane 3) or presence of GM-CSF and SCF (panel, lane 4) for an additional 24 hours. (A) Cells were harvested and stained with annexin-V and PI to examine apoptosis of each cell group. The percentage of early apoptotic cells (annexin^highPI^low) and apoptotic/dead cells (annexin^highPI^high) are shown for each cell population. (B) Cell-cycle analysis for G₀/G₁ and S+G₂/M for the corresponding cell populations. (C) Survivin expression and DNA staining for each cell population. The MCF is included in each dot plot. The area below the horizontal bar in each dot blot represents isotype control. (D) RT-PCR (35 cycles) for survivin and GAPDH mRNA for the corresponding cell populations. Negative controls indicated no amplification (not shown). (E) Western blots for survivin protein in lysates from each of the corresponding cell populations.