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1.
Fig. 4.

Fig. 4. From: Clonality of mouse and human cardiomyogenesis in vivo.

Validation of BrdU labeling. (A and B) Spectral analysis of BrdU-labeled nuclei. See text for detail. (C) Fraction of BrdU- and EdU-labeled myocytes. (D–F) BrdU (white) and EdU (green) co-localize in myocyte nuclei (arrows) at the border zone of the infarct.

Toru Hosoda, et al. Proc Natl Acad Sci U S A. 2009 October 6;106(40):17169-17174.
2.
Fig. 1.

Fig. 1. From: Clonality of mouse and human cardiomyogenesis in vivo.

Genetic tagging of CPCs in situ. (A) Cardiac niche containing seven CPCs (c-kit, white). Two CPCs are labeled by EGFP (green, arrows). One EGFP-positive myocyte (α-sarcomeric actin, α-SA, red) is visible (asterisk). (B) Scatterplots of cells co-expressing EGFP and c-kit (CPCs), EGFP and CD31 (ECs), and EGFP and Thy1.2/CD90 (fibroblasts). Forward scatter (FSC) and side scatter (SSC) are also shown. Double positive cells were back-gated and are depicted by colored dots in FSC/SSC plots.

Toru Hosoda, et al. Proc Natl Acad Sci U S A. 2009 October 6;106(40):17169-17174.
3.
Fig. 3.

Fig. 3. From: Clonality of mouse and human cardiomyogenesis in vivo.

CPCs and cardiomyogenesis. (A) FISH for EGFP viral integrants (white dot in the nucleus) found in EGFP-positive (green) myocytes (α-SA, red). Area in the rectangle is shown at higher magnification in the insets. (B) At 6 months, EGFP-labeled myocytes in the mid-portion of the LV are also BrdU-positive (white, arrowheads). One EGFP-positive BrdU-negative myocyte (asterisk) is visible. *, P < 0.05 vs. BrdU-positive myocytes. (C) Most BrdU positive (white) LV myocytes do not express EGFP (arrowheads). A few BrdU-positive EGFP-positive myocytes are present (double arrowheads). CPCs express c-kit (magenta) and are labeled by BrdU (arrows). Areas in the rectangles are shown at higher magnification in the lower panels. ***, P < 0.05 vs. Atrioventricular groove (AVG) and mid-region (Mid), respectively.

Toru Hosoda, et al. Proc Natl Acad Sci U S A. 2009 October 6;106(40):17169-17174.
4.
Fig. 6.

Fig. 6. From: Clonality of mouse and human cardiomyogenesis in vivo.

Clonal tracking of hCPCs in the regenerated myocardium. (A) Scatterplots of EGFP-positive cells isolated from the regenerated myocardium expressing c-kit, CD31, and CD146. (B) Various clones were detected in distinct cardiac cell populations from the regenerated myocardium of one treated rat (see Fig. S13 in SI Appendix for chromosome mapping and sequences). (C) Sites of integration in isolated cell populations. Some clones were common to different cell classes (same color arrowheads). Key DNA sequences are ≈80 bp shorter than the corresponding clonal bands.

Toru Hosoda, et al. Proc Natl Acad Sci U S A. 2009 October 6;106(40):17169-17174.
5.
Fig. 2.

Fig. 2. From: Clonality of mouse and human cardiomyogenesis in vivo.

Clonal marking of mouse CPCs in vivo. (A) Transcripts for α-MHC (Myh6), CD31 and procollagen (Col3a1) in myocytes (Myo), ECs and fibroblasts (Fbl). Transcript for c-kit were detected by two or three rounds of nested PCR. MC, mouse myocardium; Actb, β-actin. (B–E) FACS-sorted cardiac cells express c-kit (B, green), α-SA (C, red), CD31 (D, yellow), or procollagen (E, magenta). (F) Four distinct clones were identified in CPCs, ECs, Fbl, and Myo isolated from one mouse heart 4 months after EGFP-lentivirus injection. Bands of the same molecular weight correspond to identical sites of integration of the proviral sequence in the host genome. (see Fig. S4 in SI Appendix for clonal mapping in the mouse genome and sequences).

Toru Hosoda, et al. Proc Natl Acad Sci U S A. 2009 October 6;106(40):17169-17174.
6.
Fig. 5.

Fig. 5. From: Clonality of mouse and human cardiomyogenesis in vivo.

Viral tagging of cultured hCPCs and myocardial regeneration. (A) Genomic DNA extracted from EGFP-positive-hCPCs was subjected to three rounds of PCR. The first (1st) round of PCR resulted in a DNA smear, and the second (2nd) led to the identification of seven bands. Arrows of the same color point to corresponding bands in different lanes. After the third (3rd) round, the size of the amplicons decreased by 112 bp for the 5′-side products and 138 bp for the 3′-side products. The seven bands reflect different sites of integration of the EGFP lentivirus in the human genome. PCR products were reamplified (Re-amp) and sequenced. In this example, the reamplified products of the two bands indicated by the yellow and pink arrows are shown (see Fig. S10A in SI Appendix for sequences). (B) Multiple clones were identified in cultured hCPCs (see Fig. S10 B and C in SI Appendix for chromosome mapping and sequences). (C) Band of regenerated myocardium (arrowheads) within the infarcted region of the LV. Newly formed myocytes express α-SA (upper panel, red) and EGFP (central panel, green). Lower panel, merge of upper and central panel. Spared myocytes (*).

Toru Hosoda, et al. Proc Natl Acad Sci U S A. 2009 October 6;106(40):17169-17174.

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