Does rapamycin inhibit mTORC2?

While rapamycin acutely and directly inhibits mTORC1, only chronic administration of rapamycin can inhibit mTORC2 in some, but not all, cell lines or tissues.

Does rapamycin inhibit autophagy?

Autophagy is negatively regulated by mTOR, whose activity can be inhibited by rapamycin, a lipophilic macrolide antibiotic that is a well-established inducer of autophagy37 (Figure 2). However, its downstream targets that regulate autophagy are still unknown in mammalian cells.

How often should you fast for autophagy?

Animal studies have shown evidence of autophagy after 24 hours of fasting, which starts peaking at around 48 hours of fasting. Some studies have detected autophagy in human cultured neutrophils (the most abundant type of immune cell in the blood) after 24 hours.

How do you induce autophagy?

“Fasting is [the] most effective way to trigger autophagy,” explains Petre. “Ketosis, a diet high in fat and low in carbs brings the same benefits of fasting without fasting, like a shortcut to induce the same beneficial metabolic changes,” she adds.

What activates mTOR?

Activation of mTOR complex 1 (mTORC1) is triggered by oxidative stress, amino-acid levels and endosomal traffic to the lysosome by small GTPases such as Rab4A. In turn, mTORC1 promotes inflammation by skewing T-cell development.

How is rapamycin used in the treatment of cancer?

The drug rapamycin has important uses in oncology, cardiology, and transplantation medicine, but its clinically relevant molecular effects are not understood. When bound to FKBP12, rapamycin interacts with and inhibits the kinase activity of a multiprotein complex composed of mTOR, mLST8, and raptor (mTORC1).

How much rapamycin should I use in cell culture?

I prepared the rapamycin in DMSO (5mg in 5 mL, that is roughly 1 mM), then aliquot to 100 ul, I diluted it in serum free DMEM before use (10-fold serial dilution), then treated the cells 1h/6h. But neither my target protein nor mTOR phosphorylation is affected.

How is rapamycin used to treat Akt / PKB?

The proapoptotic and antitumor effects of rapamycin are suppressed in cells expressing an Akt/PKB mutant that is rapamycin resistant. Our work describes an unforeseen mechanism of action for rapamycin that suggests it can be used to inhibit Akt/PKB in certain cell types.

Where can I get a copy of rapamycin?

AffiliationCollaborative Innovation Center of Yangtze River Delta Region Green Pharmaceuticals, College of Pharmaceutical Sciences, Zhejiang University of Technology, Hangzhou, China Han Zhao, RolesData curation