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Fig. 3. Sf9 cells gradually regain Cry1C-sensitivity while progressing through the cell-cycle and recovering from G2-M arrest. (A) Normal (unarrested) Sf9 cells (left) and Sf9 cells released from the G2/M arrest after nocodazole had been washed off (right) and (B) G2-M-arrested cells. All cells were exposed to 300 ng/ml Cry1C for 18 hours. Dying cells with a disrupted plasma membrane were detected by staining nucleic acids with ethidium homodimer-1 (red fluorescence). (C) Time course of Cry1C-induced cell death in the three treatments shown in A and B. Most of the untreated Sf9 cells (90%) died within 6 hours of Cry1C-exposure (
). Together with cell-cycle progression in cells released from G2-M arrest, a gradual sensitivity to Cry1C was regained, detected by dead red-stained cells that started to appear after 8-9 hours of Cry1C exposure (
). Very low cell mortality was observed in G2/M-arrested cells following application of Cry1C (
).
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