Hello,
I am a Ph.D. student at the Gwangju Institute of Science and Technology in South Korea, and I have some questions regarding my immunofluorescence (IF) experiment.
In my recent experiments, I observed a bright DAPI signal in the nucleus of senescent cells following treatment with my peptide (A). Notably, this signal was absent in the untreated senescent cells. Could this bright signal be indicative of apoptotic bodies?
Additionally, I observed another distinct bright spherical structure following DAPI staining (B). Could this also represent apoptotic bodies?
Thank you.
Dear Donghoon
I can't say for sure, but the guess can be right. Even if we do not emphasize apoptotic bodies, the presence of dense chromatin, higher permeability of cell membrane and nucleus, as well as disruption of MDR activity in apoptotic cells can lead to more staining and brightness with DAPI.
Dear, Alireza Naderi Sohi, Thank you for your opinion. It's really helpful to me. And do you think A and B are equal state? Thank you for sharing your thoughts.
Hello Donghoon Kang,
Apoptosis is characterized by condensation and fragmentation of the nucleus. This leads to intense signal. So, the bright signal from DAPI can be due to apoptotic bodies. The high intensity signal is considered a sign of chromosomal condensation.
Dear Kumudini Paliwal
Thank you for your comment. It's really helpful for me. I was just wondering whether my peptide could induce apoptosis. Thank you.
Dear Donghoon Kang,
It seems to me that the peptide treatment may cause apoptosis. In order to confirm you might have to repeat this and conduct some other experiments like annexin V/PI to confirm apoptosis.
Dear Kumudini Paliwal,
Thank you for your prompt reply. I have already confirmed apoptosis using the caspase 3/7 assay, but I wasn't sure these DAPI signals were a sign of apoptosis. Thank you.