A-C. Inhibitors of cell death (zVAD-fmk and Nec-1) do not rescue the BMP2-induced decrease in cell numbers.

<p>(A) The inhibitors zVAD-fmk and Nec-1 were first tested to assess their biological activity. Jurkat A3 cells were grown overnight in 96-well plates. The following day, the cells were incubated with the inhibitors MLN (20 μM), zVAD-fmk (50 μM) and necrostatin-1 (90 μM) using the concentrations indicated. After 1 h, TNFalpha (100 ng/ml) was added, and the cells were incubated overnight. Cell viability was determined using CytoTox. (B) KMS12-BM and (C) L363 cells were incubated with the inhibitors MLN (20 μM), zVAD-fmk (50 μM) and necrostatin-1 (90 μM) at the concentrations indicated. After 1 h, BMP2 (250 nM) was added, and the cells were incubated for 96 h. Cells of the control group were left untreated. Cell number was determined by cell counting using a Coulter Counter.