Title

Advancing Metabolic Flux Analysis in the Single-Cell Era

Authors

Maggi Smart

Document Type

Article

Publication Date

Summer 2022

Keywords

JMG

JAX Location

In: Student Reports, Summer 2022, The Jackson Laboratory

Abstract

The Seahorse assay is the current, established method of measuring fuel usage in live cells in the Chang lab. The Single Cell ENergetIc metabolism by profilIng Translation inHibition (SCENITH) assay is a newly developed method of measuring cellular metabolic fuel usage that has the potential to be less expensive, less time consuming, and more adaptable to various cell types than the Seahorse assay. For this project, an EL4-EG7-OVA (EG7) (T cell lymphoma) cell line was used to validate and compare both methods using glycolysis and mitochondrial respiration (OXPHOS) inhibiting treatments. It was found that the SCENITH assay could be used to measure levels of protein translation and correlate these results to cell metabolism. Furthermore, SCENITH analysis of cell types of our lupus-prone mouse model was in line with previous Seahorse-based laboratory findings, suggesting that SCENITH is comparable to the Seahorse assay.

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