Plasmid Construct Generation for IPMK Study

The shRNA and scrambled control plasmids were from Sigma-Aldrich. The myc tagged murine WT IPMK, IPMK K129A, IPMK K129A/S235A and IPMK ΔNLS constructs were cloned to pCDH-EF1-MCS-T2A-copGFP vector (System Biosciences). The myc tagged human WT IPMK, IPMK K146A, IPMK ΔNLS and myc tagged GFP constructs were cloned to pCDH-EF1-MCS-T2A-copGFP vector (System Biosciences). The GFP tagged WT IPMK and IPMKΔNLS were cloned to pLVX-AcGFP1-N1 vector (Clontech). The flag tagged pVHL and HIF-1α genes were cloned to pLVX-AcGFP1-N1 vector (Clontech). The GST tagged GFP and PHD2 were cloned to pCDH-EF1-MCS-T2A-copGFP vector (System Biosciences). Lenti virus were generated by transfection of lenviviral vector together with pMD2.G and psPAX2 to HEK293T/17 cells^{44 (link)}.

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Fu C., Tyagi R., Chin A.C., Rojas T., Li R.J., Guha P., Bernstein I.A., Rao F., Xu R., Cha J.Y., Xu J., Snowman A.M., Semenza G.L, & Snyder S.H. (2017). Inositol Polyphosphate Multikinase Inhibits Angiogenesis via Inositol Pentakisphosphate-Induced HIF-1α Degradation. Circulation research, 122(3), 457-472.

Publication 2017

pCDH-EF1-MCS-T2A-copGFP vector pLVX-AcGFP1-N1 vector

Genes Human Murine Plasmids Pvhl Shrna Transfection vector Vector Virus

Corresponding Organization :

Other organizations : Johns Hopkins University

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Variable analysis

independent variables

Myc tagged murine WT IPMK
IPMK K129A
IPMK K129A/S235A
IPMK ΔNLS
Myc tagged human WT IPMK
IPMK K146A
IPMK ΔNLS
Myc tagged GFP
GFP tagged WT IPMK
IPMKΔNLS
Flag tagged pVHL
HIF-1α
GST tagged GFP

dependent variables

Not explicitly mentioned

control variables

Scrambled control plasmids

positive controls

Not specified

negative controls

Scrambled control plasmids

Annotations

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