IPMK-Dependent Endothelial Cell Assay

1×10⁶ wild type or IPMK KO MEFs were cultured for 24 hours; the cell-culture media were filtered (0.22 μm filter) and concentrated (ratio 1:10) by ultracentrifugal filters (10 kDa, Millipore). The concentrated cell-culture media were added to EGM2 medium (ratio 1:10) to suspend HUVECs (1×10⁵ /ml). As controls concentrated fresh DMEM was added to EGM2 medium. The HUVECs suspensions were then plated onto Matrigel. Photos were taken 4 hours later^{43 (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

ultracentrifugal filters

Cell Cell culture Matrigel

Corresponding Organization :

Other organizations : Johns Hopkins University

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Variable analysis

independent variables

Genotype (wild type or IPMK KO MEFs)

dependent variables

Tube formation by HUVECs on Matrigel (as observed in photos taken 4 hours later)

control variables

Cell culture conditions (24 hours)
Filtration (0.22 μm filter) and concentration (ratio 1:10) of cell-culture media
Ratio (1:10) of concentrated cell-culture media to EGM2 medium for HUVEC suspension
HUVEC cell density (1×10^5 /ml)

controls

Concentrated fresh DMEM added to EGM2 medium
Not explicitly mentioned

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