Background of TFEB and mTORC1
Mammalian target of rapamycin complex 1 (mTORC1) is an important regulator of cell growth. MTORC1 localizes to late endosomes/lysosomes and its dysregulation contributes to cancer, metabolic disease and diabetes. As a downstream protein of mTOR, transcription factor EB (TFEB) controls lysosomal biogenesis and cellular trafficking. Due to the essential role of lysosomes in metabolic signaling, the number and composition of lysosomes are delicately transcriptionally regulated in response to nutrient availability and other environmental factors. The MTORC1-TFEB signaling axis is a major hub that allows cells to adjust lysosome number, activity, and function according to cellular nutritional status.
Fig. 1. Describes the model of TFEB mechanical regulation proposed by MTORC1. (Martina JA, et al., 2012)
Our TFEB and mTORC1 Activity Assessment Services
MTORC1 linked to TFEB, a major regulator of lysosomal biogenesis, is a novel regulatory network, and assessing MTORC1 and TFEB activity is critical for studying the biogenesis of lysosomes. For many years, CD BioSciences has been committed to identifing TFEB and transcriptionally regulate TFEB activity to regulate the processes of lysosomal biosynthesis, autophagy, and lysosomal cytosol. Here, we offer the following services to assess MTORC1 and TFEB activity.
- Assessment of TFEB and mTORC1 Activity In Vitro
Our engineers used the following methods to assess TFEB subcellular localization and phosphorylation status to monitor mTORC1 activity.
- We monitored TFEB localization by immunofluorescence analysis using TFEB antibodies.
- We monitored the phosphorylation status of specific serine residues by WB using phosphorylation antibodies.
- We monitored the expression levels of target genes associated with lysosomal biogenesis and autophagy by quantitative PCR.
- Assessment of TFEB and mTORC1 Activity In Vivo
Our engineers used mTORC1 as an indicator of TFEB activity.
- We monitored mTORC1 signaling by immunohistochemical (IHC) techniques using antibodies that recognize the phosphorylated form of S6, an S6K substrate and an indirect substrate for mTORC1.
- We assessed the phosphorylation levels of mTORC1 substrates in tissue extracts by WB and followed mTORC1 activation status in vivo.
- We immediately show TFEB subcellular localization by directly monitoring GFP fluorescence using TFEB-GFP overexpressing cells.
- We assessed TFEB activity by WB or IHC to evaluate transcription factor E3 (TFE3) protein abundance and subcellular localization.
Why Choose Us
- Establishing animal models to further elucidate TFEB function, including lower organisms and mammals.
- Induction of TFEB activity was successfully used by us as a therapeutic strategy in several disease models.
- We use fixed cells and live imaging to regulate cellular energy metabolism by recruiting TFEB to lysosomes after activation.
- Providing theoretical support for the mechanism of activation of TFEB under conditions that induce lysosomal stress.
CD BioSciences can meet any reasonable needs of our clients, taking time and budget into consideration for you. Our aim is to be customer-centric and to provide the highest quality services to customers. Our customer service representatives are enthusiastic and trustworthy 24 hours a day, 7 days a week. If you are interested in our services, please feel free to contact us for more information or a detailed discussion.
References
- Martina JA, Chen Y, Gucek M, Puertollano R. (2012) MTORC1 functions as a transcriptional regulator of autophagy by preventing nuclear transport of TFEB. Autophagy. 8(6):903-14.
- Napolitano G, Ballabio A. (2016) TFEB at a glance[J]. Journal of cell science. 129(13): 2475-2481.