Cardiac hypertrophy in a dish: A human stem cell based model

Figures

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  Fig. 1.. Histogram plot of the FC analysis performed on the CMs used in the study. The x-axis shows the number of counts and the y-axis shows the fluorescence intensity. Blue peak represents control and red peak represents the sample. The bar signifies the interval of cTnT+ cells.

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  Fig. 2.. ICC images of control and ET-1 stimulated CMs. Cardiomyocytes stained for c-troponin T (red), F-actin (green) and Dapi (blue). The first row of images shows control CMs and the second row ET-1 stimulated CMs, both groups at 8, 24, 48, 72 and 96 h. Images were captured at 10× and the scale bars represents 100 µm.

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  Fig. 3.. RT-qPCR results from dose response experiment using CMs cultured to day 25 and day 38. The results are presented as relative fold change, normalized against the endogenous control CREBBP and unstimulated control CMs. (A) NPPA expression (B) NPPB expression (C) ACTA1 expression. Individual values are represented as black dots (day 25) and blue dots (day 38). Standard deviation (s.d.) are given as error bars (n=3). *P<0.05, **P<0.01.

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  Fig. 4.. ANP and proBNP analysis in conditioned media. Red dots represent CMs stimulated with ET-1 and black dots represent control CMs. The x-axis shows the time points and the y-axis the concentration in ng/ml. (A) Concentration of secreted ANP protein. (B) Concentration of secreted proBNP protein. s.d. is given as error bars (n=3). *P<0.05, **P<0.01, ***P<0.001.

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  Fig. 5.. Lactate concentration analysis in conditioned media. The x-axis shows the time points and the y-axis the concentration of lactate in ng/µl. Red dots represent CMs stimulated with ET-1 and black dots represent control CMs. s.d. is given as error bars (n=3). *P<0.05, **P<0.01.

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  Fig. 6.. The graph shows the cell size and the number of DEGs. The dots represent the cell volume and the squares represent DEGs. Black dots are controls and red dots represent stimulated cells. The blue dots with a black dotted line show the number of DEGs. The left y-axis shows the volume, the right y-axis the number of DEGs, and the x-axis the different time points.

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  Fig. 7.. Heatmaps of all DEGs (rows represent genes and columns represent samples). The color indicates the scaled expression values with red color representing high expression and blue representing low expression. (A) The total of 1017 DEGs (43% upregulated and 57% downregulated) at 8 h of ET-1 stimulation. (B) The total of 699 DEGs (43% upregulated and 57% downregulated) at 24 h of ET-1 stimulation. (C) The total of 155 DEGs (45% upregulated and 55% downregulated) at 48 h of ET-1 stimulation. (D) The total of 158 DEGs (68% upregulated and 32% downregulated) at 72 h of ET-1 stimulation. (E) The total of 45 DEGs (73% upregulated and 27% downregulated) at 96 h.

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  Fig. 8.. Venn diagrams showing the number of differentially expressed genes identified after ET-1 stimulation at different time of exposure. The numbers in parentheses show the total number of upregulated genes (A) and downregulated genes (B) for each exposure time. The numbers indicated in the different areas in the Venn diagrams represent the number of identified genes that overlap across the different exposure times. Five genes are identified as upregulated regardless of exposure time. The corresponding number for the downregulated genes is two. Lists of up- and downregulated genes corresponding to these Venn diagrams are shown in Table S1.

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  Fig. 9.. Heatmaps of DEGs annotated with the GO term Muscle system processes at the different time points and that are identified as significantly differentially expressed. The scale goes from green to red with the highest z-score as red and the lowest as green. The rows represent the DEGs found among the genes that are annotated with the GO-term Muscle system processes and the columns represent the samples. For all investigated time points, the majority of the identified DEGs show upregulation by ET-1 stimulation. (A) 8 h, (B) 24 h, (C) 48 h, (D) 72 h, (E) 96 h.