PMID- 27922100
OWN - NLM
STAT- MEDLINE
DCOM- 20180604
LR  - 20181113
IS  - 2045-2322 (Electronic)
IS  - 2045-2322 (Linking)
VI  - 6
DP  - 2016 Dec 6
TI  - Evaluation of candidate reference genes for RT-qPCR studies in three metabolism 
      related tissues of mice after caloric restriction.
PG  - 38513
LID - 10.1038/srep38513 [doi]
LID - 38513
AB  - Reverse transcription quantitative-polymerase chain reaction (RT-qPCR) is a 
      routine method for gene expression analysis, and reliable results depend on 
      proper normalization by stable reference genes. Caloric restriction (CR) is a 
      robust lifestyle intervention to slow aging and delay onset of age-associated 
      diseases via inducing global changes in gene expression. Reliable normalization 
      of RT-qPCR data becomes crucial in CR studies. In this study, the expression 
      stability of 12 candidate reference genes were evaluated in inguinal white 
      adipose tissue (iWAT), skeletal muscle (Sk.M) and liver of CR mice by using three 
      algorithms, geNorm, NormFinder, and Bestkeeper. Our results showed beta2m, Ppia and 
      Hmbs as the most stable genes in iWAT, Sk.M and liver, respectively. Moreover, 
      two reference genes were sufficient to normalize RT-qPCR data in each tissue and 
      the suitable pair of reference genes was beta2m-Hprt in iWAT, Ppia-Gusb in Sk.M and 
      Hmbs-beta2m in liver. By contrast, the least stable gene in iWAT or Sk.M was Gapdh, 
      and in liver was Pgk1. Furthermore, the expression of Leptin and Ppar-gamma were 
      profiled in these tissues to validate the selected reference genes. Our data 
      provided a basis for gene expression analysis in future CR studies.
FAU - Gong, Huan
AU  - Gong H
AD  - The MOH key laboratory of Geriatrics, Beijing Hospital, National Center of 
      Gerontology, Beijing, P.R. China.
FAU - Sun, Liang
AU  - Sun L
AD  - The MOH key laboratory of Geriatrics, Beijing Hospital, National Center of 
      Gerontology, Beijing, P.R. China.
FAU - Chen, Beidong
AU  - Chen B
AD  - The MOH key laboratory of Geriatrics, Beijing Hospital, National Center of 
      Gerontology, Beijing, P.R. China.
FAU - Han, Yiwen
AU  - Han Y
AD  - The MOH key laboratory of Geriatrics, Beijing Hospital, National Center of 
      Gerontology, Beijing, P.R. China.
FAU - Pang, Jing
AU  - Pang J
AD  - The MOH key laboratory of Geriatrics, Beijing Hospital, National Center of 
      Gerontology, Beijing, P.R. China.
FAU - Wu, Wei
AU  - Wu W
AD  - The MOH key laboratory of Geriatrics, Beijing Hospital, National Center of 
      Gerontology, Beijing, P.R. China.
FAU - Qi, Ruomei
AU  - Qi R
AD  - The MOH key laboratory of Geriatrics, Beijing Hospital, National Center of 
      Gerontology, Beijing, P.R. China.
FAU - Zhang, Tie-Mei
AU  - Zhang TM
AD  - The MOH key laboratory of Geriatrics, Beijing Hospital, National Center of 
      Gerontology, Beijing, P.R. China.
LA  - eng
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
DEP - 20161206
PL  - England
TA  - Sci Rep
JT  - Scientific reports
JID - 101563288
RN  - 0 (Leptin)
RN  - 0 (PPAR gamma)
RN  - 0 (RNA, Messenger)
SB  - IM
MH  - Adipose Tissue, White/metabolism
MH  - Animals
MH  - *Caloric Restriction
MH  - Gene Expression Profiling/*standards
MH  - Gene Expression Regulation
MH  - Leptin/genetics/metabolism
MH  - Male
MH  - Metabolism/*genetics
MH  - Mice, Inbred C57BL
MH  - Models, Animal
MH  - Organ Specificity/*genetics
MH  - PPAR gamma/genetics/metabolism
MH  - RNA, Messenger/genetics/metabolism
MH  - Real-Time Polymerase Chain Reaction/*standards
MH  - Reference Standards
MH  - Reproducibility of Results
MH  - Software
PMC - PMC5138604
EDAT- 2016/12/07 06:00
MHDA- 2018/06/05 06:00
PMCR- 2016/12/06
CRDT- 2016/12/07 06:00
PHST- 2016/09/13 00:00 [received]
PHST- 2016/11/09 00:00 [accepted]
PHST- 2016/12/07 06:00 [entrez]
PHST- 2016/12/07 06:00 [pubmed]
PHST- 2018/06/05 06:00 [medline]
PHST- 2016/12/06 00:00 [pmc-release]
AID - srep38513 [pii]
AID - 10.1038/srep38513 [doi]
PST - epublish
SO  - Sci Rep. 2016 Dec 6;6:38513. doi: 10.1038/srep38513.