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FIGURE 5. Effect of kinase-dead (DA) WNK2 upon NKCC1 and KCCs cotransporters activity. A, 86Rb+ uptake in oocytes injected with water, NKCC1 cRNA alone or together with WNK2-DA or WNK3-DA cRNA. Uptake was performed in isotonic conditions in the absence (open bars) or presence (closed bars) of 100 μm bumetanide. *, significantly different from the uptake observed in NKCC1 cRNA group. B, oocytes were injected with water, KCC2 cRNA alone or together with kinase-dead WNK2 or WNK3, as stated. C, oocytes were injected with water, KCC2, or KCC4 cRNA alone or together with kinase-dead WNK2 or WNK3, as stated. 86Rb+ Uptake was performed in in the presence (open bars) or absence (closed bars) of extracellular Cl−, in hypotonic (B) or isotonic (C) conditions. *, significantly different from the uptake observed in corresponding control. D, representative Western blot analysis of protein homogenates extracted from oocytes injected with wild type or kinase-dead HA-WNK2 cRNA, together with NKCC1 or KCC2 cRNA, as stated. Western blots were then performed using anti-HA monoclonal antibodies. Similar results were observed in the absence of NKCC1 cRNA injections. Image published in: Rinehart J et al. (2011) © 2011 by The American Society for Biochemistry and Molecular Biology, Inc. This image is reproduced with permission of the journal and the copyright holder. This is an open-access article distributed under the terms of the Creative Commons Attribution-NonCommercial license Larger Image Printer Friendly View

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