Immunohistochemical analysis combined with laser scanning confocal
microscopy
hiPSC-CMs were fixed in 4% paraformaldehyde, treated with Triton X-100
and blocked with 3% BSA (sigma, USA). The cells were incubated with
anti-Nav1.5 (Alomone, Israel, Lot # ASC005AN4202)
diluted in 1% BSA (1:100) overnight at 4℃, in darkness. The cell was
washed with PBS and incubated with Peroxidase-conjugated Goat
Anti-Rabbit IgG (H+L) diluted in 1% BSA (1:400) at room temperature in
the dark for 1 hour. Subsequently, DAPI was added to hiPSC-CMs, covered
with a glass slide, fixed around nail polish, and photographed by
Olympus FV1000 (Olympus, Japan). The images were analyzed with OLYMPUS
FLUOVIEW Ver.3.1b software (Olympus, Japan).