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Nirbhai Singh, Michelle Tiem, Christian Bowers, Balamurali Ambati; Role Of Heterogeneous Nuclear Ribonucleoprotein G (hnrnp-g) In Cornea. Invest. Ophthalmol. Vis. Sci. 2011;52(14):5306.
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To determine if Heterogeneous nuclear ribonucleoprotein G (hnRNP-G) regulates sFlt-1 expression in cornea
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.
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.
hnRNP-G binds to FLT-1mRNA and over expression leads to changes in sFLT-1 levels.
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