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The thymus is a vital organ of the immune system, responsible for the maturation and selection of T lymphocytes, which play a critical role in adaptive immunity. The Hedgehog (Hh) signaling pathway has been implicated in thymic differentiation and the maintenance of T-cell progenitors. However, the precise mechanisms by which Hh signaling regulates thymocyte development are not yet fully understood. This question was addressed using the ACD RNAscope™ HiPlex assay, a single-cell resolution gene expression profiling technology, to investigate the expression patterns of Hh signaling pathway components in thymic tissue.

Twelve transcripts in thymus from wildtype and Gli1 knock-out mice were analyzed using Indica Labs HALO software. The results reveal dynamic changes in the expression of Hh pathway genes during thymic differentiation, with significant changes observed between thymic compartments. Overall, these findings provide new insights into the role of Hh signaling pathway in thymic differentiation and highlight the potential of ACD’s HiPlex assay for studying complex cellular processes at single-cell resolution.

Learning Objectives

• Experience using the RNAscope™ HiPlex assay will be shared.
• Learn about creation of classifiers for the identification of thymic medulla vs thymic cortex.
• Learn about using the Indica Labs FISH v3.1.2 module in RNAscope HiPlex ISH analysis.

Speaker

Julia Jones, Ph.D. Scientific Manager, Cancer Research UK Cambridge Institute