Figure 6. Analysis of the cell cycle distribution in populations of cells with activated and nonactivated caspases.

Figure 6

Analysis of the cell cycle distribution in populations of cells with activated and nonactivated caspases. Exponentially growing HL-60 cells, untreated (A) and treated with TNF and cycloheximide for 5 h (B), were then exposed to Fam-VAD-fmk for 1 h, fixed in formaldehyde, permeabilized, incubated with RNase A and stained with PI, as described.19 Cellular green and red fluorescence was measured by flow cytometry. The increased intensity of Fam-VAD-fmk fluorescence identifies cells with activated caspases. The insets show the DNA content frequency histograms of the gated cell populations of the cells with activated (top) and nonactivated (bottom) caspases. In concordance with the published data that TNF induces apoptosis regardless of the cell cycle phase,68 activation of caspases also occurs in cells in all phases of the cycle.

From: In Situ Activation of Caspases Revealed by Affinity Labeling Their Enzymatic Sites

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