Figure 4: Functional analysis of FLNC mutants.
From: Mutations in filamin C cause a new form of familial hypertrophic cardiomyopathy
(a) H9C2 rat cardiac myoblast cells were transiently transfected with expression vectors encoding DDK-tagged wild-type FLNC or mutants p.V123A, p.A1539T, p.R2133H or p.A2430V. Soluble and insoluble protein extracts were subjected to SDS–PAGE, and filamin C or actin was detected by western blot using an anti-DDK or anti-actin antibodies, showing the presence of insoluble filamin C and actin when some mutants were expressed. (b) Subcellular distribution of ectopically expressed FLNC wild type and mutants in neonatal rat cardiac myocytes was analysed by immunofluorescence and confocal microscopy. Filamin C distribution was detected using an anti-DDK antibody, actin fibres were stained with phalloidin and nuclei were counterstained with DAPI. Ectopic expression of mutant FLNC constructs resulted in the formation of perinuclear protein aggregates not observed in wild-type FLNC.
