Fig. 4.

Characterization of β-tubulin and JP2 localization and functions in hESC-derived cardiomyocytes overexpressing SORBS2. (a)Representative confocal images of JP2 immunofluorescence staining of LVNC and control heart sections. (scale bars: 10 μm). (b) and (c) Representative immunoblot for the JP2 level in LVNC and control hearts. Quantitative analysis of the JP2 protein level in the LVNC and control hearts (n = 4 for normal hearts and n = 5 for LVNC hearts). Data shown as mean ± SEM; a Student's t-test was used for the inferential statistical analysis. (d) Co-localization of SORBS2 and β-tubulin in hESC-derived cardiomyocytes. Immunohistochemical staining of SORBS2 (red) and β-tubulin (green) proteins; the co-localized SORBS2 and β-tubulin proteins are visualized in yellow in these in hESC-derived cardiomyocytes. Nuclei were stained with DAPI (blue) The white box in the left panels represents the areas enlarged in the right panels (scale bars: 25 μm). (e) and (f) Representative images of β-tubulin immunofluorescence staining of hESC-derived CMs transfected with HBLV-h-SORBS2 and empty-virus control cells. Quantitative analysis of the expression of β-tubulin in control and SORBS2 overexpression hESC-CMs. (n = 25–30 cells per group, **p < 0.01; Student's t-test). (g) and (h) Immunoblotting to assess the β-tubulin level of hESC-derived cardiomyocytes. Quantitative analysis of the expression of β-tubulin in both its free (F) and polymerized (P) forms. Data shown as mean ± SEM; a Student's t-test was used for the inferential statistical analysis. (i) Representative images of immunofluorescence signals for JP2 and RyR2 in hESC-derived CMs. The white box in the left panels represents the areas enlarged in the right panels (scale bars: 25 μm). SORBS2-OE: hESC-CMs transfected with HBLV-h-3*flag-GFP-SORBS2 lentivirus. Control: hESC-CMs transfected with HBLV-GFP-PURO-NC lentivirus. (For interpretation of the references to color in this figure legend, the reader is referred to the web version of this article.)