• Anne von Koschembahr added an answer:
    If in addition to activation of a TNFa pathway another pathway that induces apoptosis is active, how does the cell decide to die?

    When the cell undergoes ATP depletion, apoptosis cannot occur and the cell dies by necroptosis (programmed necrosis). In TNFa -induced necroptosis, activation of Complex IIb ( Ripk1 / Ripk3 ) kills the cell without ATP depletion (I think). If in addition to activation of TNFa pathway another pathway that induces apoptosis is active, how does the cell decide to die?

    Anne von Koschembahr · University of Cincinnati

    Wouldn't it then be important to determine the various populations of cell death, in order to determine if TNF-alpha causes an apoptotic response versus necrosis? Could one say that this response may be a cell-type specific response? 

  • Dorothy C Bennett added an answer:
    Does G2/M arrest necessarily lead to cell death? How so?

    Is there a signaling pathway or a particular signal induced by arrest between S phase and mitosis (cells don't divide after synthesizing new DNA) that initiates cell death? Is this death necessarily mediated by caspases?

    Dorothy C Bennett · St George's, University of London

    DNA damage signalling can also lead to cell senescence (at least partly through CHK2-p53-p21) rather than cell death.  It is not entirely clear how cells choose one or the other, but probably with lower levels of damage signalling you would get more senescence (permanent arrest) and less death.  So cells that fail to complete S phase or mitosis can end up in a kind of G1 phase with tetraploid or subtetraploid DNA, as several groups have shown.

  • Lei Shang added an answer:
    How can I identify the cell with the staining of Annexin V (-)/PI (+) using flow cytometry?
    Annexin V (+)/PI (+) represent necrosis and later apoptosis, Annexin V (-)/PI (-) represent normal, Annexin V (+)/PI (-) represent early apoptosis. So what about Annexin V (-)/PI (+), please help me and attach your related references.
    Lei Shang · Central South University
    Maybe someone give me the answer about primary necrosis, but how to understand this kind of cell death, what the relationship between primary necrosis and necrosis??
  • Demba Sarr added an answer:
    Does anyone know the best way to induce Necroptosis in cells? TNF, TRAIL...?
    I would like to induce necroptosis in trophoblast cell line.
    Demba Sarr · University of Georgia
    Thank you Scott. I will keep you posted and sorry for the delay.
  • Lei Shang added an answer:
    How to knockdown the expression of RIP3?
    I cannot find a inhibitor of RIP3 to make the deletion. Can somebody give me some information to attenuate protein level?
    Lei Shang · Central South University
    Thanks Linkermann!! I agree with your comment.
  • Dominique Vanhecke added an answer:
    Is there a natural situation (in vivo) where intact endosomes could be released from cells?
    What happens to endosomes following cell death (apoptosis, necrosis etc).
    Dominique Vanhecke · University of Lausanne
    Thank you very much for your input.
    But I was wondering whether in addition to these microvesicles there can also be intact organelles, in this case intact endosomes be released under some circumstances. A big difference is that in contrast to microvesicles the endosomes would be so-called "inside-out" with respect to the orientation of membrane bound proteins.

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