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. The pMØ were infected in vitro with L. monocytogenes or vehicle for 4 h. Mice were injected with L. monocytogenes (or vehicle) and euthanized 24 h post-injection. The RGs came from a multispecies primer set, from the literature or designed here. The RG ranking relied on GeNorm, NormFinder, BestKeeper, Delta-CT and 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 but modestly 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.

中文翻译：

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单核细胞增生李斯特菌感染小鼠模型基因表达谱的参考基因。

RT-qPCR 剖析基于转录的过程，但需要参考基因 (RG) 来进行数据标准化。本研究展望了感染单核细胞增生李斯特菌的小鼠巨噬细胞 (pMØ) 和脾脏的 RG。 pMØ 在体外被L 感染。单核细胞增多症 或载体 4 小时。给小鼠注射L。单核细胞增生李斯特菌（或媒介物）并在注射后 24 小时安乐死。 RG 来自多物种引物组，来自文献或此处设计。 RG 排名依赖于 GeNorm、NormFinder、BestKeeper、Delta-CT 和 RefFinder。 B2m-H3f3a-Ppia 是 pMØ 最稳定的 RG，尽管 RG 索引对细胞因子相对表达的估计进行了微调。 Actβ-Ubc-Ppia 是对脾脏最好的 RG，但影响不大细胞因子的相对表达。因此，小鼠模型为L。单核细胞增生李斯特菌需要特定环境的 RG 进行 RT-qPCR，从而强化了其对准确基因表达谱分析的重要贡献。