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Rupp et al. Genomic DNA. Exon 6. Exon 9. Exon 10. cDNA from mRNA. mutation. 302bp. Supplementary Figure 1: Structure of the human troponin T gene.
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Rupp et al. Genomic DNA Exon 6 Exon 9 Exon 10 cDNA from mRNA mutation 302bp Supplementary Figure 1: Structure of the human troponin T gene Parts of genomic DNA and cDNA from Troponin T (TnT) are shown. The mutation site (G>A) is located at position 8,728 in exon 9. Primers of human TnT were designed to span from exon 6 to exon 10. The length of the analyzed cDNA is 302bp.
Rupp et al. Supplementary Figure 2: Immunostaining of differentiated CPC human cell marker merge a-sarcomeric actinin CM-Dil a-sarcomeric actinin hNA Immunohistochemistry of co-cultured circulating progenitor cells (CPC) and rat cardiomyocytes after 6 days. The co-culture assay was performed as previously described (7,9). Patient-derived CM-DiL-labelled CPC (upper panel) or human nuclear antigen (hNA) positive cells (lower panel) express α-sarcomeric actinin (green) and display differentiated human cardiomyocytes.
Rupp et al. #1 #2 #3 Donor A: Sequence of 7 clones #4 #5 #6 #7 Donor B Supplementary Figure 3: Wildtyp and mutated sequence of human TnT 7 clones derived from RNA of co-cultured CPC of donor A (green box) were sequenced. The wild type human TnT sequence was found in 4 clones. The mutated TnT sequence was found in 3 clones (blue filled box). Sequence of a clone isolated from control (donor B) is shown in red box.
Rupp et al., h-TnT hr-GAPDH human heart CPC CM Fixed CM + CPC h-Nkx2.5 h-MLC-2V h-ANP GAPDH CM Fixed CM + CPC Supplementary Figure 4: RT-PCR for cardiospecific markers after co-culture of CPC on fixed cardiomyocytes RT-PCR for human TnT in CPC cultivated for 6 days with previously paraformaldehyde- fixed cardiomyocytes (1 %, 15 min). Human TnT is expressed in human heart and in differentiated CPC after co-culture of CPC and cardiomyocytes (CM). Human TnT is not expressed in rat cardiomyocytes and in CPC before co-culture. RT-PCR for additional cardiac marker genes as indicated in CPC after co-culture with paraformaldehyde-fixed rat cardiomyocytes for 6 days is shown in the lower panel. Rat cardiomyocytes served as negative control. Human and rat GADPH was used as loading control.