FIG. 4.
The loss of plasma membrane integrity is associated with a large VAC in cells. Fibroblasts of beige mice (Lystbg) or control mice (C57BL/6) (A) and C57BL/6 fibroblasts of different passages (B) were treated with or without TNF-α plus CHX for 10 h. Half of each sample was analyzed by LysoTracker staining, and the other half was analyzed by PI exclusion plus FSC. The relative value of the VAC was determined by the intensity of LysoTracker staining. The results represent means ± standard errors (n = 3). The percentages of R1 and R2 cells are shown. (C) Jurkat cells were cotransfected with the BAX expression vector or an empty vector and a green fluorescent protein (GFP) expression vector. The cells were left untreated or treated with zVAD (50 μM) for 12 h after transfection. GFP expression was used to identify transfected cells by using a fluorescence-activated cell sorter. The relative value of the VAC in transfected cells was measured by staining the cells with LysoTracker at 12, 18, 24, and 30 h after transfection. The viability of cells was analyzed 30 h after transfection by PI exclusion plus FSC. Percentages of R1 and R2 are shown.