Good morning all,
I am using an inducible system with doxycycline for my cells, I cannot see a knockdown in my western blot. My target gene is involved in cytokinesis.
I am currently trying different timepoints and concentration (1ug/mL to 3ug/mL). Is it possible that the cells that I see dead in my plate are the ones that have the knockdown?
Thank you very much :(!
Best,
Cédric
Hi Cédric,
possibly this is the case. For example it is known that doxycycline inhibits Nf-kB and thereby can induce apoptosis.
https://www.oncotarget.com/article/12488/text/
If the gene you want to knockdown is also directly affected by doxycycline, using this approach for knockdown might be problematic...
Best wishes,
Christina
Is there any chance that knocking down the gene itself could be lethal?