I am using MDA-MB-231 breast cancer cell line. after 24 hr treatment many vacuoles is formed but Apoptosis is not occurring even after 48 hr of treatment. Why this vacuoles are formed. Is it linked with Autophagy ?
Go J Yoshida
It is generally difficult to determine the presence of autophagosomes with optical microscope unless you will stain cells with LC3-II antibody.
1 votes
1 thanks
Rola Jaafar
whats the treatment?? try staurosporine control and see how they look..
0 votes
0 thanks
Ammar Kurdi
Negative ctrl: untreated, positive ctrl: treat cells with 1 uM rapamycin or EBSS medium(starvation) Add bafilomycin A1 (50-100 nM) 24 hours later, do a Western Blot for LC3 (recommended to use the nanotools antibody) and p62 (cell signaling antibod) -offcourse beta actin also-
More LC3II and p62 ---> autophagy is involved. I recommend using different concentrations of your treatment to see if the reaction is dose dependent.
Success!
1 votes
0 thanks
Sandeep Rajput
What method are you using to determine apoptosis? Can u put some pictures here.
0 votes
0 thanks
- Badges
- Science topic
More Jayant Dewangan's questions See All
Similar questions and discussions