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Fig. 4. Binding of 14-3-3 and CaM to Kir/Gem. (A) 14-3-3 binding. Cells were cotransfected with cDNAs for wt or mutated Myc-Kir/Gem and GST–14-3-3, GST–14-3-3 K49E or GST–14-3-3 dimerization mutant. (a) GST–14-3-3 proteins were precipitated and associated Myc-Kir/Gem was detected by western blot using Myc antibody. (b and c) Cell lysates were blotted with Myc (b) or GST (c) antibodies to monitor Kir/Gem and GST–14-3-3 expression levels. (B) CaM binding. (a) Cells were transfected with cDNAs for wt or mutated Myc-Kir/Gem. Cell homogenates were incubated with CaM beads and bound Myc-Kir/Gem detected by western blot using a Myc antibody. (b) Cell lysates were blotted with Myc antibodies to monitor Kir/Gem expression levels.





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