西亚试剂：Figure 1 - Lineage tracing of EndMT in Tie1Cre;R26RstoplacZ

a) Gene constructs present in Tie1Cre;R26RstoplacZ mice. (b) An example of an unbanded control heart, in which lacZ expression was detected only in vessels (blue staining, arrows; eosin counterstain, red). (c) Representative sections of a banded heart, where lacZ expression was detected throughout the fibrotic tissue (arrows). The arrow in the right picture points to a blue cell directly adjacent to a vessel. (d) Confocal immunofluorescence double-labeling in banded Tie1Cre;R26RstoplacZ hearts with antibodies to -gal (red) and FSP1 (green). Arrows, colocalization of -gal and FSP1 expression involving both microvessels (left) and arterioles (right). (e) Normal unbanded control hearts and hearts of banded FSP1–GFP mice were stained for GFP (green) and CD31 (red). Confocal microscopy revealed GFP expression in few fibroblasts (GFP⁺CD31^- cells) in normal hearts (left, arrow points to a GFP⁺CD31^- cell). In banded hearts, GFP⁺ endothelial cells co-expressed GFP and CD31 (right; arrows point to GFP⁺CD31⁺ cells). Nuclei were counterstained with TOPRO-3 (blue). (f) Single cells from unbanded and banded Tie1Cre;R26RstoplacZ mice were FACS sorted according to lacZ and CD31 expression. Top, negative and positive controls (from a heart from Rosa26 mice, in which all cells express lacZ). LacZ activity was detected with FDG-green (x axis); CD31 protein was labeled with PE-red (y axis). In normal hearts (bottom left), lacZ⁺CD31⁺ cells were most prominent (gate R4). This population was decreased in fibrotic hearts (bottom right) in favor of lacZ⁺CD31^- cells (gate R6). (g) Cells from gates R4 and R6 of two banded Tie1Cre;R26RstoplacZ hearts were sorted and compared by real-time PCR. Expression of mRNAs encoding the mesenchymal markers -SMA, FSP1, DDR-2 and collagen I 1 (Col1A1) in the lacZ⁺CD31^- (R6) cell population of banded hearts is shown compared with their expression in the lacZ⁺CD31⁺ (R4) cell population. Scale bars, 20 m.

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