I have perform Hypoxic treatment on AML-12 cells for 12hrs and there is significant up regulation of RIP3 in western blot and no expression of cleaved casp3. I did flow cytometry with PI annexin and in Q2 theres more necrotic cells in hypoxic samples. I used gsk inhibitor to inhibit rip3 and it showed decrease in flow cytometry. So is it apoptosis or necroptosis. Also i havnot been able to see TNFA changes in treated cells. If its necroptosis, which other pathway or cell death initiator it can be?

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