DZNep-induced cell death is not necroptosis.

<p>(<b>a</b>) Responsive 8226 and resistant LP1 cells were either treated with vehicle or DZNep (1 µM for 24 h) then examined by transmission electronic microscopy. (<b>b</b>) Responsive 8226 and resistant LP1 cells were either treated with vehicle or DZNep (1 µM for 24 h). The perimeter (in µm), the area (in µm²) and the circularity (in arbitrary unit) of vehicle-(in white) or DZNep-treated (in black) LP1 and 8226 cells were determined by image analysis as described in the method section. (<b>c</b>) L363 cells were treated with vehicle, DZNep 1 µM for 24 h, or hydrogen peroxide (H₂O₂), incubated with CM-H₂DCFDA before FACS analysis. At least, 2×10⁴ events were gated. The percentage of stained cells (<i>i.e</i>. cells producing ROS) is indicated on the graph. (<b>d</b>) Responsive 8226 and resistant LP1 cell lines were treated for 6 or 24 h with DZNep 1 µM. The transcriptional expression of <i>TXNIP</i> was determined by qRT-PCR using the ΔΔC_t method with <i>RPLP0</i> as internal standard. The fold change was calculated as 2^{−ΔΔ<i>Ct</i>}. Indicated values corresponded to the mean ± SD from at least three independent experiments done with two distinct RNA preparations.