Abstract
RT-qPCR dissects transcription-based processes but requires reference genes (RGs) for data normalization. This study prospected RGs for mouse macrophages (pMØ) and spleen infected with Listeria monocytogenes. ThepMØ were infected in vitro with L. monocytogenes or vehicle for 4h. Mice were injected with L. monocytogenes (or vehicle) and euthanized 24h post-injection. TheRGs came from a multispecies primer set, from theliterature or designed here. The RG ranking relied on GeNorm, NormFinder, BestKeeper, Delta-CTand RefFinder. B2m-H3f3a-Ppia were the most stable RGs for pMØ, albeit RG indexes fine-tuned estimations of cytokine relative expression. Actβ-Ubc-Ppia were the best RGs for spleen butmodestly impacted the cytokine relative expression. Hence, mouse models of L. monocytogenes require context-specific RGs for RT-qPCR, thus reinforcing its paramount contribution to accurate gene expression profiling.
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