Abstract
Purpose: :
To determine if Heterogeneous nuclear ribonucleoprotein G (hnRNP-G) regulates sFlt-1 expression in cornea
Methods: :
Full length (hnRNP-G) was cloned into pCMV vector. pCMV- hnRNP-G and empty pCMV was transected into HUVEC cells. After 72 hrs cell were harvested and Real Time PCRs were performed for soluble FLT1 (sFLT-1) and membrane FLT-1 (mFLT-1) gene expression. Real time PCR was performed using primers designed against N- terminal region present in soluble NRP-1 and C-terminal end present in membrane NRP-1 only. RNA-binding protein immunoprecipitation (RIP) was performed using anti-hnRNP-G antibody. Bound RNA was extracted and analyzed.
Results: :
pCMVhnRNP-G over expression in HUVEC lead to 2.5 fold increase in sFLT-1 while there was no significant change in mFLT-1 levels. hnRNP-G was found to bind to FLT-1 mRNA.
Conclusions: :
hnRNP-G binds to FLT-1mRNA and over expression leads to changes in sFLT-1 levels.
Keywords: cornea: endothelium • cornea: basic science • gene/expression