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Fig. 5. An intact CNBD is specifically involved in trafficking of HERG. (A) Immunoblot performed on HEK cells expressing cDNAs encoding HERGwt and HERG{Delta}750-870. (B) HL-1 cardiac, PC12 neuronal and Cos7 kidney cells transfected with the indicated constructs and subjected to immunoblotting. (C) Immunocytochemical analyses of cells permeabilized with Triton X-100. Staining was performed with an anti-HA antibody followed by incubation with a secondary antibody conjugated with Oregon Green. Punctate staining of the HERGwt channel is absent from cells transfected with HERG{Delta}750-870. (D) Cells expressing the HERG{Delta}750-870 construct were treated with PK followed by immunoblotting. (top) The membrane was blotted with an anti-HA antibody, stripped and reprobed for CASK (bottom). Signal intensities before and after PK treatment were measured and quantified. (E) Immunoblot of cell extracts expressing HERG{Delta}26-135 and HERG{Delta}750-870 mutant channels. Membrane was probed with an anti-HERG antibody.





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