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. 2011 Aug 10;6(8):e22946. doi: 10.1371/journal.pone.0022946

Figure 5. AAV-induced cell death is independent of necroptosis.

Figure 5

(a) DAPI staining showing micronucleated cells among infected U2OSp53DD cells treated with necrostatin-1 (necro). Necrostatin-1 treatment alone did not have a significant effect on the viability of the cells. Images were acquired using the 10× objective. Arrows indicate micronucleated cells. The inserts show cells at a higher magnification. Bar: 230 µm. (b) The experiment described in (a) was replicated and the average percentage of micronucleated cells was calculated in infected cells with or without necrostatin-1 treatment. Error bars represent standard deviations from two independent experiments. Un: untreated. (c) PI staining and FACS analysis showing that AAV-induced cell death in U2OSp53DD cells is not sensitive to necrostatin-1 (x-axis: DNA content; y-axis: cell count). Necrostatin-1 treatment alone did not have an effect on the cell cycle. (d) Control FACS experiment to test the effectiveness of necrostatin-1. NIH3T3 cells were treated with TNFα and zVAD-fmk to induce necroptosis, which was then successfully inhibited by necrostatin-1. These experiments were performed with the DAKO flow cytometer (see Materials and Methods) and so have a slightly different presentation from the other FACS analyses.