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. 2009 Aug 24;29(21):5872–5888. doi: 10.1128/MCB.00112-09

FIG. 1.

FIG. 1.

Characterization of GMX1778 cytotoxicity. (A) Kinetics of NAD+ depletion, ATP depletion, and cell lysis. Cellular NAD+ levels, measured by LC/MS, and ATP depletion data are from one experiment, while the cell lysis data are from an independent experiment. Data are presented as means ± standard deviations. (B) Clonogenicity, annexin V reactivity, and cell lysis (propidium iodide [PI] permeability) of IM-9 cells after 72 h of exposure to GMX1778 (25 nM). Clonogenicity was assessed in soft agar by counting colonies, and data are presented as percentages (± standard errors of the means) of colonies compared to levels of DMSO-treated cells. Annexin V reactivity and PI permeability were assessed by flow cytometric analyses of 10,000 events. (C) Caspase 3 activity and cleavage in IM-9 cells treated with GMX1778. Caspase 3 activity was measured in 25 μg of cell extract. Inset, Western blot of the accumulation of cleaved caspase 3 in 25 μg of extracts from cells treated with GMX1778. The panels are from a single Western blot, with intervening lanes removed.