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. 2017 Apr 26;8:502. doi: 10.3389/fimmu.2017.00502

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Copyright © 2017 Kunkl, Porciello, Mastrogiovanni, Capuano, Lucantoni, Moretti, Persson, Galandrini, Buzzetti and Tuosto.

This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) or licensor are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.

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ISA-2011B inhibits exogenously expressed PIP5Kα lipid-kinase activity. (A) Jurkat cells were transfected and cultured for 24 h with 20 µg control empty vector (Vec) or 20 µg HA-PIP5Kα and then treated for further 6 h with DMSO or 25 µM ISA-2011B. PIP5Kα kinase assays performed on anti-HA IP (upper panel). HA-PIP5Kα content was analyzed by western blotting (lower panel). (B) Fold inductions were quantified by densitometric analysis and normalized to HA-PIP5Kα levels. Data are expressed as fold induction over the basal level of cells transfected with empty vector and treated with DMSO. Bars show the mean ± SD of two experiments. Asterisks (***) indicate P < 0.001 calculated by Student’s t-test, compared with cells transfected with HA-PIP5Kα and treated with DMSO.