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Fig. 4. The effect of cell confluence on sensitivity to osmotic shock. NEB-1 (wild-type keratin) cells were grown to different degrees of confluence (40%, 80% and 100%), before subjecting them to hypoosmotic shock; metabolic activity was measured at 1, 3 and 24 hours of recovery (CellTiter 96, Promega). At all time points, cultures at 80% confluence showed the greatest sensitivity to osmotic shock.