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. 2020 Mar 11;11:1315. doi: 10.1038/s41467-020-14958-x

Fig. 2. Injection of Sema7a results in increased MIRI and PNC formation.

Fig. 2

Mice were injected with either recombinant semaphorin 7a (rmSema7a) or Fc control (rmIgG2A Fc) and then subjected to 1 h of ischemia followed by 2 h reperfusion. Samples were taken after 1 or 120 min reperfusion. a Representative TTC-stained slices of myocardial tissue showing infarcted area (blue/dark = retrograde Evans blue staining; red and white = AAR, white = infarcted tissue; 120 min) with b systematic evaluation of infarct sizes and corresponding troponin I plasma levels (120 min, n = 6/group). c Representative histological sections (scale bar 100 µm) of WT animals injected with either IgG Fc control or rmSema7a (120 min) and d number of PNCs counted in myocardial tissue sections of AAR in animals (120 min, n = 9/group). e Representative flow-cytometric plots of PNCs in the blood of sham animals, rmSema7a- or IgG Fc control–treated mice, showing GPIb (CD42b) and P-selectin (CD62P) after 1 and 120 min reperfusion. Systematic evaluation of flow-cytometric expression of mean fluorescence intensity (MFI) for f GPIb (CD42b, n = 6;6;4;4;6) and g P-selectin (CD62P, n = 6;3;5;3;7) expression. h Systematic evaluation of PNCs in % by flow cytometry in the blood of animals injected with rmSema7a or Fc control at 1 and 120 min (n = 6;4;4;4;5). i Representative flow-cytometric plots of PNCs in the AAR showing the presence of GPIb (CD42b) and P-selectin (CD62P) after 1 and 120 min reperfusion. Systematic evaluation of flow-cytometric expression of MFI for j GPIb (CD42b, n = 5;5;5;4;6) and k P-selectin (CD62P, n = 6;4;5;3;5) and l systematic evaluation of PNCs in % by flow cytometry in the AAR of animals injected with rmSema7a or Fc control at 1 and 120 min (n = 5;4;5;5;5). Comparisons in b and d were analyzed by unpaired two-tailed Student’s t-tests (data are mean ± SD). For f–h, j–l we used log transformation of data to conform normality. For log-transformed data, unpaired two-tailed Student’s t-tests were performed on the log values and results are displayed as geometric means and their 95% confidence intervals. *p < 0.05, **p < 0.01, and ***p < 0.001 as indicated.