TGFß1-mediated expression and alternative splicing of Fibronectin Extra Domain A in human podocyte culture.
Cell Mol Biol (Noisy-le-grand)
; 64(3): 17-24, 2018 Feb 28.
Article
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| MEDLINE
| ID: mdl-29506626
ABSTRACT
Alternative splicing is a fundamental phenomenon to build protein diversity in health and diseases. Extra Domain A+ Fibronectin (EDA+Fn) is an alternatively spliced form of fibronectin protein present in the extra cellular matrix (ECM) in renal fibrosis. Podocytes are spectacular cell type and play a key role in filtration and synthesise ECM proteins in renal physiology and pathology. TGFß1 is a strong stimulator of ECM proteins in renal injury. In this study, we have investigated alternative splicing of EDA+ Fn in human podocytes in response to TGFß1. We have performed western blotting and immunofluorescence to characterise the expression of the EDA+Fn protein, real-time PCR for RNA expression and RT-PCR to look for alternative splicing of EDA+Fn in conditionally immortalised human podocytes culture.We used TGFß1 as a stimulator and SB431542 and SRPIN340 for inhibitory studies. In this work, for the first time we have demonstrated in human podocytes culture EDA+Fn is expressed in the basal condition and TGFß1 2.5ng/ml induced the Fn mRNA and EDA+Fn protein expression demonstrated by real-time PCR, western blotting and immunofluorescence. TGFß1 2.5ng/ml induced the alternative splicing of EDA+Fn shown by conventional RT-PCR. Studies with ALK5 inhibitor SB431542 and SRPIN340 show that TGFß1 induced alternative splicing of EDA+Fn was by the ALK5 receptor and the SR proteins. In human podocytes culture, alternative splicing of EDA+Fn occurs at basal conditions and TGFß1 further induced the alternative splicing of EDA+Fn via ALK5 receptor activation and SR proteins. This is the first evidence of basal and TGFß1 mediated alternative splicing of EDA+Fn in human podocytes culture.
Palabras clave
Texto completo:
1
Colección:
01-internacional
Banco de datos:
MEDLINE
Asunto principal:
Fibronectinas
/
Empalme Alternativo
/
Podocitos
/
Factor de Crecimiento Transformador beta1
Límite:
Humans
Idioma:
En
Revista:
Cell Mol Biol (Noisy-le-grand)
Asunto de la revista:
BIOLOGIA MOLECULAR
Año:
2018
Tipo del documento:
Article
País de afiliación:
Reino Unido