Biomedical Engineering Reference
In-Depth Information
Fig. 1.
Characterization of HEK-KWGF cells.
a
Western blot analysis of non-transfected HEK-
293 cells (HEK), Kir2.1-GFP expressing HEK-293 cells (HEK-KWGF) and rat neonatal
cardiomyocytes (NCMs). Kir2.1-GFP signal (GFP) was only observed in HEK-KWGF cells.
Connexin43 is expressed at low levels in HEK and HEK-KWGF cells, and at high levels in
NCM. Cadherin adherence junction proteins (pan-Cadherin) are expressed in all three cell types
at similar levels. Total protein staining is used as loading control.
b
GFP fluorescence
microscopy displays strong plasma membrane localization of Kir2.1-GFP.
c
Current traces of
HEK-KWGF cells characteristic for
I
K1
. Scale bars—horizontal = 100 ms,vertical = 1,000 pA.
d
Current-voltage relationship of HEK-KWGF cells displaying
I
K1
characteristics
of high levels of functional
I
K1
channels in these cells, displaying an I-V curve
characteristic for Kir2.1 channels (Fig. 1c, d). Current densities were -58.7 ±
6.6 pA/pF at -100 mV, which is approximately 6-15 fold higher than in
isolated neonatal (approximately -4 pA/pF) [23] and adult murine heart cells
