Figure 5
From: Distinct functional defect of three novel Brugada syndrome related cardiac sodium channel mutations
Representative western blotting of WT, p.I848fs. p.R965C, and p.1876insM SCN5A channels from 4 independent experiments. HEK293T transfected with WT or mutant plasmid were firstly treated with cell impermeable biotinylation reagent and total proteins were extracted from these cells. After immunoprecipitated with anti-myc antibody, which recognized the epitope added on all SCN5A constructs, the immunoprecipitated complex were subject to western blotting and reacted with HRP conjugated streptavidin. All four SCN5A constructs displayed bands by western blotting, indicating they were all subject to biotinylation reaction. This suggested that they all reached cytoplasmic membrane. Note that p.I848fs produced a truncated protein with a smaller molecular weigh than others.