start-ver=1.4
cd-journal=joma
no-vol=29
cd-vols=
no-issue=2
article-no=
start-page=279
end-page=284
dt-received=
dt-revised=
dt-accepted=
dt-pub-year=2014
dt-pub=201402
dt-online=
en-article=
kn-article=
en-subject=
kn-subject=
en-title=
kn-title=Functional analysis of NPHS1 mutations in Japanese patients
en-subtitle=
kn-subtitle=
en-abstract=
kn-abstract=Background: Many mutations in the NPHS1 gene were detected among patients with congenital nephrotic syndrome. Functional analysis of those mutations was done with a stable-expression cell line. Nevertheless, establishing such a cell line is time-consuming. Methods and Results: We established an easier method using automatic counting software for functional analysis with transient-transfection cells rather than a stable-expression cell line. We demonstrated maltrafficking to the plasma membrane of abnormal nephrin for immunostaining on transient-expression cells by comparison without Triton X (detecting proteins on the cell membrane only) and with Triton X (detecting proteins both on the cell membrane and inside the cell cytoplasm). We obtained relevant results with data obtained previously using a stable-expression cell line. Furthermore, we conducted functional analysis of NPHS1 mutations in Japanese patients with congenital nephrotic syndrome using this simple method, which revealed that all pathogenic mutations impaired trafficking to the protein plasma membrane. 

Conclusions: Functional analysis using transient-expression cells with automatic counting software was useful to demonstrate maltrafficking to the plasma membrane of a protein. All pathogenic mutations detected in Japanese patients impaired trafficking to the protein plasma membrane.
en-copyright=
kn-copyright=

en-aut-name=MiyaiTakayuki
en-aut-sei=Miyai
en-aut-mei=Takayuki
kn-aut-name=
kn-aut-sei=
kn-aut-mei=
aut-affil-num=1
ORCID=

en-aut-name=AyaKunihiko
en-aut-sei=Aya
en-aut-mei=Kunihiko
kn-aut-name=
kn-aut-sei=
kn-aut-mei=
aut-affil-num=2
ORCID=

en-aut-name=TakaiwaMasanori
en-aut-sei=Takaiwa
en-aut-mei=Masanori
kn-aut-name=
kn-aut-sei=
kn-aut-mei=
aut-affil-num=3
ORCID=

en-aut-name=YanKunimasa
en-aut-sei=Yan
en-aut-mei=Kunimasa
kn-aut-name=
kn-aut-sei=
kn-aut-mei=
aut-affil-num=4
ORCID=

en-aut-name=SadoYoshikazu
en-aut-sei=Sado
en-aut-mei=Yoshikazu
kn-aut-name=
kn-aut-sei=
kn-aut-mei=
aut-affil-num=5
ORCID=

en-aut-name=TanakaHiroyuki
en-aut-sei=Tanaka
en-aut-mei=Hiroyuki
kn-aut-name=
kn-aut-sei=
kn-aut-mei=
aut-affil-num=6
ORCID=

en-aut-name=MorishimaTsuneo
en-aut-sei=Morishima
en-aut-mei=Tsuneo
kn-aut-name=
kn-aut-sei=
kn-aut-mei=
aut-affil-num=7
ORCID=

affil-num=1
en-affil=
kn-affil=Okayama Univ, Grad Sch Med Dent & Pharmaceut Sci, Dept Pediat

affil-num=2
en-affil=
kn-affil=Okayama Univ, Grad Sch Med Dent & Pharmaceut Sci, Dept Pediat

affil-num=3
en-affil=
kn-affil=Okayama Univ, Grad Sch Med Dent & Pharmaceut Sci, Dept Pediat

affil-num=4
en-affil=
kn-affil=Kyorin Univ, Sch Med, Dept Pediat

affil-num=5
en-affil=
kn-affil=Shigei Med Res Inst, Div Immunol

affil-num=6
en-affil=
kn-affil=Okayama Saiseikai Gen Hosp, Dept Pediat

affil-num=7
en-affil=
kn-affil=Okayama Univ, Grad Sch Med Dent & Pharmaceut Sci, Dept Pediat
en-keyword=Congenital nephrotic syndrome
kn-keyword=Congenital nephrotic syndrome
en-keyword=Nephrin
kn-keyword=Nephrin
en-keyword=Podocin
kn-keyword=Podocin
en-keyword=Trafficking
kn-keyword=Trafficking
en-keyword=Transient transfection
kn-keyword=Transient transfection
END