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01.09.2009 | Original Research Paper | Ausgabe 9/2009

Inflammation Research 9/2009

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

Zeitschrift:
Inflammation Research > Ausgabe 9/2009
Autoren:
Congshan Jiang, Liesu Meng, Wenhua Zhu, Muhammad Shahzad, Xudong Yang, Shemin Lu
Wichtige Hinweise
Responsible Editor: J. A. Di Battista.

Electronic supplementary material

The online version of this article (doi:10.​1007/​s00011-009-0027-5) contains supplementary material, which is available to authorized users.
C. Jiang and L. Meng contributed equally to the work.

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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Zusatzmaterial
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_MOESM1_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_MOESM2_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_MOESM3_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)
11_2009_27_MOESM4_ESM.tif
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