pS6 immunoprecipitation RNA-seq of mouse olfactory epithelium in response to single odorants, binary mixtures, and complex fragrances

Phosphorylation of ribosomal protein S6 (pS6) serves as a molecular marker of neuronal activation by external stimuli. In this study, olfactory epithelium tissues were collected from mice exposed to single odorants (acetophenone, decanal, octanal, and cis-3-hexenol), binary mixtures (acetophenone–decanal and octanal–cis-3-hexenol), or complex fragrances (floral, mint, and floral–mint combination). To selectively capture transcripts undergoing active translation in odor-activated cells, pS6-associated ribosome complexes were isolated from tissue lysates through immunoprecipitation. The enriched mRNAs were subsequently purified and subjected to RNA sequencing, enabling transcriptomic profiling specifically within odor-activated neuronal populations, particularly olfactory sensory neurons. This approach provides a targeted view of gene expression programs induced by diverse odor stimulation paradigms in the heterogeneous olfactory epithelium.