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. 2019 Nov 12;13:498. doi: 10.3389/fncel.2019.00498

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Copyright © 2019 Bergmann, Garcia de Paco, Rissiek, Menzel, Dubberke, Hua, Rassendren, Ulmann and Koch-Nolte.

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) and the copyright owner(s) 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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Flow cytometry analysis of mouse P2X4 expression using Nodu 246. (A) Cell surface and intracellular expression of mouse P2X4 by P2X4-transfected and non-transfected HEK cells was assessed by flow cytometry after sequential incubation of cells with rat mAb Nodu 246 followed by a secondary PE-conjugated anti-rat-IgG antibody (2nd antibody), in non-permeabilized or permeabilized cells. Similar experiments were performed with BMDM obtained from WT and P2X4^−/− mice. Cells were either non-permeabilized (B) or permeabilized (C). Note that permeabilization greatly enhanced the detection of mouse P2X4 in BMDM, further supporting the intracellular localization of the native receptor.