314 Chapter 12 Figure 5. Increased fibroblast activation expression and protein expression in PLN-R14del spheroids. (A) Representative immunofluorescent imaging of alpha-actinin (green). (B) Representative immunofluorescent imaging of alpha-actinin (green) co-stained with Ki67 (red). (C) Representative immunofluorescent imaging of alpha-actinin (green) co-stained with Pro-Collagen 1 (red). Hoechst is used to stain the nuclei (Blue). Scale bar: 500 μm. Immunofluorescent quantification of hCSs α-Actinin expression (D), proliferation marker Ki67 (E), and Pro-Collagen 1 (F) per nuclei. ** P< 0.01 vs. CTR group by Student’s unpaired T-test. Each dot represents one individual spheroid and 2-3 biological and 3-10 technical replicates per condition were included in this analysis. (G) qPCR analysis of various genes involved in fibroblast activation or cardiomyocyte function in PLN-R14del hCSs vs. controls. Each dot represents the expression of one sample containing 20 hCSs, normalized for average control spheroid expression. (H) Heat map and hierarchical clustering of gene expression data. Log 2-fold changes of gene expression are represented and the color scale indicates the level of change (blue, low; red, high). The hierarchical clustering was generated using the Euclidean distance correlation metric based on the square root of the sum of the square differences (https://software.broadinstitute.org/morpheus).
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