Figure 3. Analysis of translation of wt and mutant α subunits and of assembly with β subunits. Oocytes were not injected or injected with RNAs encoding ENaC carrying a FLAG epitope on the β subunit, αβFγ, αS589LβFγ, αS589VβFγ, αS589WβFγ, or βFγ, as indicated. (A) Western blot immunostaining of solubilized wt and mutant ENaC subunits. Solubilized proteins were subjected to SDS-PAGE, and αENaC was visualized on Western blots using an anti–rat αENaC antibody (May et al. 1997). Molecular markers are indicated. (B) Western blot immunostaining for coimmunoprecipitation of wt and mutant ENaC under nondenaturing conditions. Nondenaturing coimmunoprecipitation was performed with an anti-FLAG antibody. Immunoprecipitates were subjected to SDS-PAGE and αENaC was visualized on Western blots using an anti–rat αENaC antibody. Specificity of the anti–rat αENaC antibody is demonstrated by a control in which no αENaC RNA was injected (βFγ).
Image published in: Kellenberger S et al. (2001)
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