Supporting Figures

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Supporting Figure 6
Supporting Figure 7




Supporting Figure 6

Fig. 6. Effect of the catalytically inactive WNK3-D294A (with no lysine = K, WNK) on K-Cl cotransporter (KCC)4 activity under hypotonic conditions, in oocytes injected with differing amounts of KCC4 cRNA. ⁸⁶Rb⁺ uptake was measured in Xenopus laevis oocytes microinjected with KCC4 cRNA alone or together with WNK3-D294A cRNA. KCC4 cRNA was injected at two different concentrations, 0.05 and 0.2 mg/ml, that result in total injection of 2.5 and 10 ng of cRNA per oocyte. Uptakes were performed in hypotonic medium (110 mOsm/kg) in the presence (open bars) or absence (filled bars) of extracellular Cl^– in the uptake medium. Each bar represents the mean ± SEM of 10 oocytes. *, P < 0.001 vs. control without WNK3-D294A.

Supporting Figure 7

Fig. 7. Effect of protein phosphatases inhibitors on K-Cl cotransport induced by cell swelling. ⁸⁶Rb⁺ uptake in oocytes injected with KCC1 (A), KCC2 (B), KCC3 (C), and KCC4 (D). All groups were injected with cRNA concentration at 0.2 mg/ml, except for KCC1, which was injected at 0.4 mg/ml. Uptakes were performed in hypotonic conditions in oocytes that were exposed to Cl^–-containing medium (open bars) or Cl^–-depleted medium (filled bars). Each bar represents the mean ± SEM of 20 oocytes from two different frogs. Protein phosphatase inhibitors used were as follows: calyculin A (at 100 nM), okadaic acid (at 1 nM), and cyclosporine A (at 25 mM). *, significantly different from the uptake observed in the corresponding control (absence of protein phosphatase inhibitors, P < 0.01).