Figure 5. IF analysis of myosin isoform... | Rockefeller University Press

Figure 5.

$IF analysis of myosin isoform expression in single hESC-CMs replated at different times. (A–E) hESC-CMs grown for (A) 34, (B) 42, (C) 45, (D) 48, and (E) 55 d on laminin-coated glass coverslips were detached, replated on laminin-coated glass coverslips, grown for the indicated number of days (+d), and compared with non-replated controls (d). Myosin isoform expression was analyzed by single-cell IF using specific antibodies against α- and β-MyHC. The fractions of cells in the different categories are shown as a percentage of the total number of cells analyzed (n, set to 100%) for each time point (see Materials and methods). One coverslip from one differentiation each. (F) Changes in sarcomeric gene expression (log2 fold change) of hESC-CMs after replating compared with 37 d old non-replated controls. hESC-CMs grown for 35 d on laminin-coated glass coverslips were detached, replated on laminin-coated glass coverslips and cultivated for two more days. mRNA expression was analyzed using RNA-Seq data. Changes in mRNA expression of indicated myosin heavy chain (MYH), myosin light chain (MYL), and troponin (TNN) isoform genes are presented as log2 fold change. The dashed line at ±0.585 indicates a ±1.5-fold change; n = 3 culture dishes from three differentiations.$

IF analysis of myosin isoform expression in single hESC-CMs replated at different times. (A–E) hESC-CMs grown for (A) 34, (B) 42, (C) 45, (D) 48, and (E) 55 d on laminin-coated glass coverslips were detached, replated on laminin-coated glass coverslips, grown for the indicated number of days (+d), and compared with non-replated controls (d). Myosin isoform expression was analyzed by single-cell IF using specific antibodies against α- and β-MyHC. The fractions of cells in the different categories are shown as a percentage of the total number of cells analyzed (n, set to 100%) for each time point (see Materials and methods). One coverslip from one differentiation each. (F) Changes in sarcomeric gene expression (log2 fold change) of hESC-CMs after replating compared with 37 d old non-replated controls. hESC-CMs grown for 35 d on laminin-coated glass coverslips were detached, replated on laminin-coated glass coverslips and cultivated for two more days. mRNA expression was analyzed using RNA-Seq data. Changes in mRNA expression of indicated myosin heavy chain (MYH), myosin light chain (MYL), and troponin (TNN) isoform genes are presented as log2 fold change. The dashed line at ±0.585 indicates a ±1.5-fold change; n = 3 culture dishes from three differentiations.