Ischemic cardiac stromal fibroblast-derived protein mediators in the infarcted myocardium and transcriptomic profiling at single cell resolution

Figures

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  Fig. 1. The level of secreted proteins of ischemia challenged CF using mass spectrometry. A Pie diagram showing the proportion of proteins based on the FC expression in the ISC vs Control groups, B Scatter diagram of newly upregulated proteins in ISC group which were absent in the control, C Scatter diagram showing the FC distribution of proteins expressed in both ISC and control groups. D Scatter diagram of newly downregulated proteins in ISC group which were present exclusively in the control. E Pie diagram showing the proportion of proteins based on the FC expression in the ISC/R vs Control groups, F Scatter diagram of newly upregulated proteins in ISC/R group which were absent in the control, G Scatter diagram showing the FC distribution of proteins expressed in both ISC/R and control groups. H Scatter diagram of newly downregulated proteins in ISC/R group which were present exclusively in the control. I Pie diagram showing the proportion of proteins based on the FC expression in the ISC vs ISC/R groups, J Scatter diagram of newly upregulated proteins in ISC group which were absent in the ISC/R, K Scatter diagram showing the FC distribution of proteins expressed in both ISC and ISC/R…

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  Fig. 2. Representative images for the immunofluorescence analysis for the expression of Cofilin 1 in the LV (A) (N = 3) and the quantification of protein expression of Cofilin 1 in LV tissues calculated from MFI (B). C Quantification of Cofilin 1 gene expression in CF (N = 8) following the treatment with ISC and ISC/R using qRT-PCR relative to the housekeeping gene GAPDH normalized to the control and presented as Log2 FC. Representative images for the immunofluorescence and quantification for the expression of Cofilin 1 in the CF (N = 3) following the treatment with ISC and ISC/R (D and E). Representative image of Western blot and quantification showing the expression status of Cofilin 1 in the ISC and ISC/R groups compared to the control in CF (N = 4) (F and G). Single cell genomics of Cofilin 1 + CF showing the (H) (split and combined views of Control, ISC and ISC/R groups of t-SNE plot) showing the distribution of cells within all clusters based on the global expression of 4601 genes. Violin plot (I) (FC 5 and 5.5) indicating the alteration of genes in the 3 clusters. J Scatter plot FC expression indicating the alteration of Cluster 1 genes reflecting the contrasting phenotype…

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  Fig. 3. Representative images for the immunofluorescence analysis for the expression of CRSP2 in the LV (A) (N = 3) and the quantification of protein expression of CRSP2 in LV tissues calculated from MFI (B). C Quantification of CRSP2 gene expression in CF (N = 4) following the treatment with ISC and ISC/R using qRT-PCR relative to the housekeeping gene GAPDH normalized to the control. Representative images for the immunofluorescence and quantification for the expression of CRSP2 in the CF (N = 3) following the treatment with ISC and ISC/R (D and E). Single cell genomics of CRSP2 + CF showing the (F) (split and combined views of Control, ISC and ISC/R groups of t-SNE plot) showing the distribution of cells within all clusters based on the global expression of 6821 genes. Violin plot (G) (FC > 2) indicating the alteration of genes in the 6 clusters. J Scatter plot FC expression indicating the alteration of Cluster 1 genes reflecting the expression of the key signature genes. (**** P < 0.0001, and unlabeled groups represent NS)

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  Fig. 4. Representative images for the immunofluorescence analysis for the expression of HSP27 in the LV (A) (N = 3) and the quantification of protein expression of HSP27 in LV tissues calculated from MFI (B). C Quantification of HSP27 gene expression in CF (N = 3) following the treatment with ISC and ISC/R using qRT-PCR relative to the housekeeping gene GAPDH normalized to the control. Representative images for the immunofluorescence and quantification for the expression of HSP27 in the CF (N = 3) following the treatment with ISC and ISC/R (D and E). Representative image of Western blot and quantification showing the expression status of HSP27 in the ISC and ISC/R groups compared to the control in CF (N = 4) (F and G). Single cell genomics of HSP27 + CF showing the (H) (split and combined views of Control, ISC and ISC/R groups of t-SNE plot) showing the distribution of cells within all clusters based on the global expression of 6821 genes. Violin plot (I) (FC > 5) indicating the alteration of genes in the 2 clusters. J Scatter plot FC expression indicating the alteration of Cluster 1 and 2 genes reflecting the contrasting level of expression of the key signature genes. (* P < 0.05, ** P…

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  Fig. 5. Representative images for the immunofluorescence analysis for the expression of IL8 in the LV (A) (N = 3) and the quantification of protein expression of IL8 in LV tissues calculated from MFI (B). C Quantification of IL8 gene expression in CF (N = 4) following the treatment with ISC and ISC/R using qRT-PCR relative to the housekeeping gene GAPDH normalized to the control. Representative images for the immunofluorescence and quantification for the expression of HSP27 in the CF (N = 3) following the treatment with ISC and ISC/R (D and E). Representative image of Western blot and quantification showing the expression status of IL8 in the ISC and ISC/R groups compared to the control in CF (N = 3) (F and G). Single cell genomics of IL8 + CF showing the (H) (split and combined views of Control, ISC and ISC/R groups of t-SNE plot) showing the distribution of cells within all clusters based on the local expression of 2654 genes. Violin plot (I) (FC > 2) indicating the alteration of genes in the 2 clusters. J Scatter plot FC expression indicating the alteration of Cluster 1 genes reflecting the expression of key signature genes. (* P < 0.05, *** P < 0.001, **** P < 0.0001, and unlabeled…

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  Fig. 6. Representative images for the immunofluorescence analysis for the expression of HSP90 in the LV (N = 3) (A) and the quantification of protein expression of HSP90 in LV tissues calculated from MFI (B). C Quantification of HSP90 gene expression in CF (N = 4) following the treatment with ISC and ISC/R using qRT-PCR. Representative images for the immunofluorescence and quantification for the expression of HSP90 in the CF (N = 3) (D and E) (N = 3) following the treatment with ISC and ISC/R. F Representative image of Western blot and (G) quantification showing the expression status of HSP90 in the ISC and ISC/R groups compared to the control in CF (N = 4). Single cell genomics of HSP90 + CF showing the (H) (split and combined views of Control, ISC and ISC/R groups of t-SNE plot) showing the distribution of cells within all clusters based on the local expression of 4272 genes. Violin plot (I) (FC > 5) indicating the alteration of genes in the 2 clusters. J Scatter plot FC expression indicating the alteration of Cluster 1 genes reflecting the expression of key signature genes. (* P < 0.05, ** P < 0.01, **** P < 0.0001, and unlabeled groups represent NS)

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  Fig. 7. Representative images for the immunofluorescence analysis for the expression of Nrf2 in the LV (N = 3) (A) tissues and (B) the quantification of protein expression of Nrf2 in LV tissues calculated from MFI. C Quantification of Nrf2 gene expression in CF (N = 4) following the treatment with ISC and ISC/R using qRT-PCR. Representative images for the immunofluorescence and quantification for the expression of Nrf2 in the CF (N = 3) (D and E) following the treatment with ISC and ISC/R. F Representative image of Western blot and (G) quantification showing the expression status of Nrf2 in the ISC and ISC/R groups compared to the control in CF (N = 4). Single cell genomics of Nrf2 + CF showing the (H) (split and combined views of Control, ISC and ISC/R groups of t-SNE plot) showing the distribution of cells within all clusters based on the local expression of 5817 genes. Violin plot (I) (FC > 4) indicating the alteration of genes in the 2 clusters. J Scatter plot FC expression indicating the alteration of Cluster 1 genes reflecting the expression of key signature genes. (* P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001, and unlabeled groups represent NS)

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  Fig. 8. Representative images for the immunofluorescence analysis for the expression of Troponin-I (Trop I) in the LV (N = 3) (A) tissues and the quantification of protein expression of Troponin-I in LV tissues calculated from MFI (B). C Quantification of Troponin-I gene expression in CF (N = 4) following the treatment with ISC and ISC/R using qRT-PCR. Representative images for the immunofluorescence and quantification for the expression of Troponin-I in the CF (N = 3) (D and E) following the treatment with ISC and ISC/R. F Representative image of Western blot and (G) quantification showing the expression status of Troponin-I in the ISC and ISC/R groups compared to the control in CF (N = 4) (**** P < 0.0001, *** P < 0.001, ** P < 0.01, * P < 0.05, and unlabeled groups represent NS)