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. 2020 Nov 27;9:e61050. doi: 10.7554/eLife.61050

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© 2020, Li et al

This article is distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use and redistribution provided that the original author and source are credited.

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Figure 8. — (A) Representative immunostainings of ZI+BZ of WT1:ROSA hearts treated or not with BNP 10 days after surgery and stained with DAPI (nuclei in blue) and antibody against GFP protein (green). Hearts represented in full in the small inserts. The orange rectangles delimited the enlarged area below. (B) Representative immunostainings of WT1:ROSA hearts treated or not with BNP 10 days after MI and stained with DAPI (nuclei in blue) and antibodies against CD31 protein (red) and GFP (green). White arrows represented GFP⁺ CD31⁺ cells, that is endothelial cells originating from WT1⁺ cells. (C) High magnification of a part of the ZI+BZ of infarcted BNP-treated hearts where WT1⁺ cells contributed to the vessel formation (orange rectangle). (D) GFP⁺ cell number per mg of cardiac tissue 3 or 10 days after surgery, determined by flow cytometry analysis. Results in BNP-treated hearts related to those obtained in saline-treated hearts. E and F. Flow cytometry analysis on isolated NMCs stained with antibodies against CD31 and GFP. (E) Percentages of CD31⁺ cells originating from WT1⁺ precursor cells (GFP⁺CD31⁺ cells). The percentages of GFP⁺ cells determined among the selected CD31⁺ cells. (F) Percentages of differentiating WT1⁺ cells into CD31⁺ cells. The percentages of CD31⁺ cells determined among the selected GFP⁺ cells. (D:) 3 days after surgery: MI: n = 4, MI+BNP: n = 6. (D–F:) 10 days after surgery: MI: n = 6, MI+BNP: n = 7 different mice. Individual values are represented and the means ± SEM are represented in red. *p≤0.05 only for groups with n ≥ 6. No difference of variance between groups.

Figure 8—figure supplement 1. — (A) Representative immunostainings of ZI+BZ of WT1:ROSA hearts treated or not with BNP 10 days after surgery and stained with DAPI (nuclei in blue) and antibody against GFP protein (green). Hearts represented in full in the small inserts. The orange rectangles delimited the enlarged area below. (B) Representative immunostainings of WT1:ROSA hearts treated or not with BNP 10 days after MI and stained with DAPI (nuclei in blue) and antibodies against CD31 protein (red) and GFP (green). White arrows represented GFP⁺ CD31⁺ cells, that is endothelial cells originating from WT1⁺ cells. (C) High magnification of a part of the ZI+BZ of infarcted BNP-treated hearts where WT1⁺ cells contributed to the vessel formation (orange rectangle). (D) GFP⁺ cell number per mg of cardiac tissue 3 or 10 days after surgery, determined by flow cytometry analysis. Results in BNP-treated hearts related to those obtained in saline-treated hearts. E and F. Flow cytometry analysis on isolated NMCs stained with antibodies against CD31 and GFP. (E) Percentages of CD31⁺ cells originating from WT1⁺ precursor cells (GFP⁺CD31⁺ cells). The percentages of GFP⁺ cells determined among the selected CD31⁺ cells. (F) Percentages of differentiating WT1⁺ cells into CD31⁺ cells. The percentages of CD31⁺ cells determined among the selected GFP⁺ cells. (D:) 3 days after surgery: MI: n = 4, MI+BNP: n = 6. (D–F:) 10 days after surgery: MI: n = 6, MI+BNP: n = 7 different mice. Individual values are represented and the means ± SEM are represented in red. *p≤0.05 only for groups with n ≥ 6. No difference of variance between groups.