The α/β 1 karyopherin complex regulates hERG1 NP nuclear transport in hiPSC-CMs. (A) hiPSC-CMs following an 8hr incubation in either vehicle (Top), 40 µM importazole (Middle), or 2.5 µM ivermectin (Bottom), and stained for hERG1 with the C-Term antibody (magenta) and DAPI (cyan). (B) Box and whisker plots depicting nuclear fluorescence intensity relative to cytoplasmic fluorescence intensity (F Nucleus /F Cytoplasm ) for the C-Term antibody. Importazole and ivermectin significantly reduced the nuclear enrichment of the C-Term antibody. Data were compared using a Kruskal-Wallis ANOVA. * indicates P < 0.05. **** indicates P < 0.0001

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... and importazole, respectively (44,45). We incubated hiPSC-CMs in 40 µM importazole for 8 h and then measured the relative nuclear and cytoplasmic distribution of hERG1 using the C-terminal antibody as described above (Fig. 2). 40 µM importazole significantly reduced the relative intensity of the nuclear hERG1 signal compared to vehicle controls ( Fig. 3), indicating that nuclear transport of the hERG1 NP is karyopherin-β1 dependent. To test if hERG1 NP trafficking is also dependent upon the α karyopherin, we blocked incubated cells with 2.5 µM ivermectin for 8 h. Like importazole, ivermectin significantly reduced the relative intensity of the nuclear hERG1 signal, compared to vehicle ...

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... 3), indicating that nuclear transport of the hERG1 NP is karyopherin-β1 dependent. To test if hERG1 NP trafficking is also dependent upon the α karyopherin, we blocked incubated cells with 2.5 µM ivermectin for 8 h. Like importazole, ivermectin significantly reduced the relative intensity of the nuclear hERG1 signal, compared to vehicle controls (Fig. 3). These data indicate that the classical nuclear transport pathway mediates hERG1 NP localization in the nucleus. The intracellular distribution of the P-Loop antibody was unaffected by cardiomyocyte maturation. Paired groups in "C" were compared using a Wilcoxon matched-pairs test. Means in "D" and "E" were compared using a ...