14-3-3θ is a Binding Partner of Rat Eag1 Potassium Channels
Figure 2
Isoform specificity of 14-3-3 binding with rEag1 N- and C-termini.
(A) GST pull-down assay of cell lysates from HEK293T cells transfected with various myc-tagged 14-3-3 isoforms. (Left panel) Coomassie blue staining of the GST proteins. (Right panel) Immunoblotting of pull-down products with the anti-myc antibody. Input volume shown at the bottom corresponds to 5% of the total cell lysates for pull-down. (B) Quantification of the pull-down efficiency of different 14-3-3 isoforms. The protein band intensities of individual myc-14-3-3 isoforms affinity precipitated by GST-N207 or GST-C0 in (A) were divided by those of cognate total inputs, thereby minimizing the potential bias conferred by the variation in protein expression among different 14-3-3 isoforms. Densitometric scans of immunoblots were obtained from three independent experiments.