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Housekeeping gene stability in pristane-induced arthritis and antigen-induced pulmonary inflammation of rats

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Abstract

Objective and design

Aim of this study is to explore the expression stability of 12 commonly used housekeeping genes in inflammation condition.

Materials and methods

Two rat models, pristane-induced arthritis (PIA) and antigen-induced pulmonary inflammation (AIPI) were established and reverse transcription-PCR was performed from rat spleens and inguinal lymph nodes.

Results

The reference gene expression fluctuated in the immune-associated tissues and 18S rRNA expression from the various inflammatory conditions was the most stably in all examined gene expression. Besides, peptidylprolyl isomerase A and beta-glucuronidase in PIA spleens, beta-actin and GAPDH in PIA inguinal lymph nodes, and peptidylprolyl isomerase A and beta-actin in AIPI spleens, expressed more stably than others. These results were further confirmed by expression study of TNF-alpha in PIA rat inguinal lymph nodes.

Conclusion

We suggest that the stable gene mentioned above or geometric mean of three best references in each experimental condition should be used as the reference and the variable ones should be avoided in similar study designs.

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Acknowledgments

We should express gratitude to Mr. Fujun Zhang, Ms.Yan Han and Qilan Ning for their expertise and assistance in the experiments. The work was supported by grants from the National Natural Science Foundation of China (30630058, 30571725 and 30801027) and Shaanxi Province International Cooperation Foundation of China (2007-KW-06).

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Authors and Affiliations

  1. Department of Genetics and Molecular Biology, Xi’an Jiaotong University School of Medicine, Post Box 81, Western Yanda Road 76, Xi’an, Shaanxi, 710061, People’s Republic of China

    Congshan Jiang, Liesu Meng, Wenhua Zhu, Muhammad Shahzad, Xudong Yang & Shemin Lu

  2. Key Laboratory of Environment and Genes Related to Diseases (Xi’an Jiaotong University), Ministry of Education, Xi’an, China

    Congshan Jiang, Liesu Meng, Wenhua Zhu, Muhammad Shahzad, Xudong Yang & Shemin Lu

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  1. Congshan Jiang
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  2. Liesu Meng
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  3. Wenhua Zhu
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  4. Muhammad Shahzad
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Corresponding author

Correspondence to Shemin Lu.

Additional information

Responsible Editor: J. A. Di Battista.

C. Jiang and L. Meng contributed equally to the work.

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11_2009_27_MOESM1_ESM.tif

Fig. A. Expression variation of twelve chosen housekeeping genes in inguinal lymph nodes of PIA rats mRNA expression levels of 18S (a), ACTB (b),GAPDH (c), HPRT (d), GUSB (e), HMBS (f), PPIA (g), RPL13A (h), ARBP (i), B2M (j),TBP (k) and SDHA (l) in spleens from individual PIA rat are plotted. Rats are divided into three groups named as control, PIA d6 and PIA d26 (n=8 for each group). “p” value is calculated by Kruskal-Wallis test. (TIFF 136 kb)

11_2009_27_MOESM2_ESM.tif

Fig. B. Expression variation of twelve chosen housekeeping genes in spleens of AIPI rats mRNA expression levels of 18S (a), ACTB (b),GAPDH (c), HPRT (d), GUSB (e), HMBS (f), PPIA (g), RPL13A (h), ARBP (i), B2M (j),TBP (k) and SDHA (l) in spleens from individual PIA rat are plotted. Rats are divided into three groups named as control (n=6), disease (n=6) and medicine (n=5). “p” value is calculated by Kruskal-Wallis test. (TIFF 134 kb)

11_2009_27_MOESM3_ESM.tif

Fig. C. Expression stability rank determined by geNorm software of 12 chosen housekeeping genes in PIA rat inguinal lymph nodes The degression of M value is represented with a solid line and two kinds of dash lines, meanwhile the rank results are shown with normal, bold italic and underlined characters for control, PIA d6 and PIA d26 groups, respectively. (TIFF 54 kb)

11_2009_27_MOESM4_ESM.tif

Fig. D. Expression stability rank determined by geNorm software of 12 chosen housekeeping genes in AIPI rat spleens The degression of M value is represented with a solid line and two kinds of dash lines; meanwhile the rank results are shown with normal, bold italic and underlined characters for control, disease and medicine groups, respectively. (TIFF 52 kb)

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Jiang, C., Meng, L., Zhu, W. et al. Housekeeping gene stability in pristane-induced arthritis and antigen-induced pulmonary inflammation of rats. Inflamm. Res. 58, 601–609 (2009). https://doi.org/10.1007/s00011-009-0027-5

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  • DOI: https://doi.org/10.1007/s00011-009-0027-5

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