ER/Autophagic Stress Assay

Endoplasmic reticulum (ER) stress plays a crucial role in the pathology of diverse diseases. Furthermore, autophagy is tightly linked to ER stress to counteract possible damaging effects resulting from impaired protein folding. Therefore, there is great interest in administering this pathway to treat clinical diseases. Creative Biolabs has established a complete technical platform in order to provide professional and customized ER/autophagic stress assay services to customers all over the world.

Overview of ER Stress

ER, known as the protein folding checkpoint, is responsible for the correct folding of proteins, and its ability to keep protein folding in balance has important implications for the function of cells. This balance can be disrupted by multiple conditions, resulting in ER stress, such as calcium fluctuations, hypoxia, inflammatory cytokines, viral infections, and nutrient deficiencies. ER stress is implicated in various diseases, such as cancer, neurodegenerative diseases, and metabolic diseases. Therefore, uncovering the mechanisms of ER stress involved in these diseases may provide potential drug targets for the treatment of these diseases.

Fig.1 ER stress signaling pathways. (Ren, et al., 2021)

ER Stress and Autophagy

To ensure the proper functioning of ER, eukaryotic cells develop an unfolded protein response (UPR) repair pathway. Evidence suggests that ER stress and autophagy are closely linked since that i) the key ER stress pathway UPR stimulates autophagy; ii) ER stress induces autophagic responses through promoting the assembly of pre-autophagosome structures. Conversely, autophagy has been reported to ameliorate ER stress by eliminating accumulated misfolded proteins. It is increasingly clear that ER stress leads to the induction of autophagy, which is a multistep catabolic process that degrades large protein aggregates and damaged organelles in autophagosomes.

ER/Autophagic Stress Assay

Creative Biolabs has mastered multiple techniques to perform ER/autophagic stress assays. In our assay, we mainly relied on flow cytometry assays to explore ER stress and autophagy. We employed flow cytometry to measure reticulophagy, combining with analysis of LC3-II and cell cycle analysis of cellular misfolded proteins. What's more, we determined the extent of misfolded protein aggregate formation in a cell cycle-dependent manner using a probe that fluoresces when bound to misfolded proteins in fixed cells. Therefore, we could investigate the properties of the observed drug-induced ER stress by detecting changes in ER mass, misfolded proteins, and the autophagy marker LC3-II.

With our advanced technology and professional scientists, Creative Biolabs focuses on providing customers with comprehensive and cost-effective ER/autophagic stress assay services. If you have any difficulties with your ER stress and autophagy project, please do not hesitate to contact us for professional advice and assistance.

Reference

1. Ren, H.; et al. The cross-links of endoplasmic reticulum stress, autophagy, and neurodegeneration in parkinson’s disease. Frontiers in Aging Neuroscience. 2021, 13: 691881.