PMID- 25658431
OWN - NLM
STAT- MEDLINE
DCOM- 20160108
LR  - 20220129
IS  - 1932-6203 (Electronic)
IS  - 1932-6203 (Linking)
VI  - 10
IP  - 2
DP  - 2015
TI  - Sub-cellular localisation studies may spuriously detect the Yes-associated 
      protein, YAP, in nucleoli leading to potentially invalid conclusions of its 
      function.
PG  - e0114813
LID - 10.1371/journal.pone.0114813 [doi]
LID - e0114813
AB  - The Yes-associated protein (YAP) is a potent transcriptional co-activator that 
      functions as a nuclear effector of the Hippo signaling pathway. YAP is oncogenic 
      and its activity is linked to its cellular abundance and nuclear localisation. 
      Activation of the Hippo pathway restricts YAP nuclear entry via its 
      phosphorylation by Lats kinases and consequent cytoplasmic retention bound to 
      14-3-3 proteins. We examined YAP expression in liver progenitor cells (LPCs) and 
      surprisingly found that transformed LPCs did not show an increase in YAP 
      abundance compared to the non-transformed LPCs from which they were derived. We 
      then sought to ascertain whether nuclear YAP was more abundant in transformed 
      LPCs. We used an antibody that we confirmed was specific for YAP by 
      immunoblotting to determine YAP's sub-cellular localisation by 
      immunofluorescence. This antibody showed diffuse staining for YAP within the 
      cytosol and nuclei, but, noticeably, it showed intense staining of the nucleoli 
      of LPCs. This staining was non-specific, as shRNA treatment of cells abolished 
      YAP expression to undetectable levels by Western blot yet the nucleolar staining 
      remained. Similar spurious YAP nucleolar staining was also seen in mouse 
      embryonic fibroblasts and mouse liver tissue, indicating that this antibody is 
      unsuitable for immunological applications to determine YAP sub-cellular 
      localisation in mouse cells or tissues. Interestingly nucleolar staining was not 
      evident in D645 cells suggesting the antibody may be suitable for use in human 
      cells. Given the large body of published work on YAP in recent years, many of 
      which utilise this antibody, this study raises concerns regarding its use for 
      determining sub-cellular localisation. From a broader perspective, it serves as a 
      timely reminder of the need to perform appropriate controls to ensure the 
      validity of published data.
FAU - Finch, Megan L
AU  - Finch ML
AD  - School of Chemistry and Biochemistry, University of Western Australia, Crawley, 
      Western Australia, 6009, Australia.
FAU - Passman, Adam M
AU  - Passman AM
AD  - School of Chemistry and Biochemistry, University of Western Australia, Crawley, 
      Western Australia, 6009, Australia.
FAU - Strauss, Robyn P
AU  - Strauss RP
AD  - School of Chemistry and Biochemistry, University of Western Australia, Crawley, 
      Western Australia, 6009, Australia.
FAU - Yeoh, George C
AU  - Yeoh GC
AD  - School of Chemistry and Biochemistry, University of Western Australia, Crawley, 
      Western Australia, 6009, Australia; Harry Perkins Institute of Medical Research, 
      QEII Medical Centre, Nedlands and Centre for Medical Research, the University of 
      Western Australia, Crawley, Western Australia, 6009, Australia.
FAU - Callus, Bernard A
AU  - Callus BA
AD  - School of Chemistry and Biochemistry, University of Western Australia, Crawley, 
      Western Australia, 6009, Australia.
LA  - eng
GR  - 09-0084/AICR_/Worldwide Cancer Research/United Kingdom
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
DEP - 20150206
PL  - United States
TA  - PLoS One
JT  - PloS one
JID - 101285081
RN  - 0 (Adaptor Proteins, Signal Transducing)
RN  - 0 (Cell Cycle Proteins)
RN  - 0 (Phosphoproteins)
RN  - 0 (Transcription Factors)
RN  - 0 (YAP-Signaling Proteins)
RN  - 0 (YAP1 protein, human)
RN  - 0 (Yap1 protein, mouse)
SB  - IM
MH  - Adaptor Proteins, Signal Transducing/genetics/*metabolism
MH  - Animals
MH  - Cell Cycle Proteins
MH  - Cell Line
MH  - Cell Nucleolus/genetics/*metabolism
MH  - Humans
MH  - Mice
MH  - Phosphoproteins/genetics/*metabolism
MH  - Protein Transport/physiology
MH  - Transcription Factors
MH  - YAP-Signaling Proteins
PMC - PMC4320119
COIS- Competing Interests: The financial support from the funders does not alter the 
      authors' adherence to PLOS ONE policies on sharing data and materials.
EDAT- 2015/02/07 06:00
MHDA- 2016/01/09 06:00
PMCR- 2015/02/06
CRDT- 2015/02/07 06:00
PHST- 2014/07/16 00:00 [received]
PHST- 2014/11/04 00:00 [accepted]
PHST- 2015/02/07 06:00 [entrez]
PHST- 2015/02/07 06:00 [pubmed]
PHST- 2016/01/09 06:00 [medline]
PHST- 2015/02/06 00:00 [pmc-release]
AID - PONE-D-14-31786 [pii]
AID - 10.1371/journal.pone.0114813 [doi]
PST - epublish
SO  - PLoS One. 2015 Feb 6;10(2):e0114813. doi: 10.1371/journal.pone.0114813. 
      eCollection 2015.