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Evaluation of suitable reference genes for gene expression studies in porcine PBMCs in response to LPS and LTA.


ABSTRACT: BACKGROUND: As an in vitro model porcine peripheral blood mononuclear cells (PBMCs) is frequently used as for immunogenetic research with the stimulation of bacterial antigens. To investigate the immunocompetence of PBMCs for recognition of Gram-positive and Gram-negative bacteria and in order to dissect the pathogenesis of diseases, gene expression assay is most commonly used. The gene expressions are required to normalize for reference genes which have tremendous effect on the results of expression study. The reference genes should be stably expressed between different cells under a variety of experimental conditions, but recent influx of data showed that expression stability of reference genes are varied under different experimental conditions. But data regarding the expression stability of reference genes in porcine PBMCs are limited. Therefore, this study was aimed to know whether the expression stability of commonly used reference genes in PBMCs is affected by various bacterial antigens under different experimental conditions in pigs. RESULTS: The mRNA expression stability of nine commonly used reference genes (B2M, BLM, GAPDH, HPRT1, PPIA, RPL4, SDHA, TBP and YWHAZ) was determined by RT-qPCR in PBMCs that were stimulated by LPS and LTA in vitro as well as cells un-stimulated control and non-cultured were also consider for this experiment. mRNA expression levels of all genes were found to be affected by the type of stimulation and duration of the stimulation (P?

SUBMITTER: Cinar MU 

PROVIDER: S-EPMC3584940 | biostudies-literature | 2013

REPOSITORIES: biostudies-literature

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Evaluation of suitable reference genes for gene expression studies in porcine PBMCs in response to LPS and LTA.

Cinar Mehmet Ulas MU   Islam Mohammad Ariful MA   Pröll Maren M   Kocamis Hakan H   Tholen Ernst E   Tesfaye Dawit D   Looft Christian C   Schellander Karl K   Uddin Muhammad Jasim MJ  

BMC research notes 20130208


<h4>Background</h4>As an in vitro model porcine peripheral blood mononuclear cells (PBMCs) is frequently used as for immunogenetic research with the stimulation of bacterial antigens. To investigate the immunocompetence of PBMCs for recognition of Gram-positive and Gram-negative bacteria and in order to dissect the pathogenesis of diseases, gene expression assay is most commonly used. The gene expressions are required to normalize for reference genes which have tremendous effect on the results o  ...[more]

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