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Macrophages are closely associated with protruding CMs at the border zone.A Immunostaining of GFP and F-actin in thick sections from Tg(mpeg1.1:NTR-YFP) ventricles at 10 dpci. White boxes denote the zoomed image from the wound border zone in (i) and (ii). B Still image from time-lapse imaging of Tg(mpeg1:EGFP); Tg(myl7:mKATE-CAAX) ventricular sections at 10 dpci. Yellow arrowheads point to mpeg1:EGFP+ cells containing myl7:mKATE-CAAX fluorescence. C Immunostaining of GFP and mScarlet in Tg(mpeg1:EGFP); Tg(myl7:lck-mScarlet) ventricles at 7 and 10 dpci. Yellow boxes denote the zoomed image from the wound border zone. D Quantification of the number of mpeg1:EGFP+ cells 50 μm proximal and distal to the wound border at 4 (n = 5 ventricles), 7 (n = 5 ventricles), 10 (n = 6 ventricles), and 14 dpci (n = 4 ventricles). Data are presented as mean ± SD. P-values were calculated using one-way ANOVA with Tukey’s multiple comparisons test. Source data are presented in the Source Data file. E Immunostaining of GFP, Tnfa, Cxcr4b, and F-actin in Tg(mpeg1:EGFP) ventricles at 10 dpci. Yellow box denotes the zoomed image from the wound border zone. F Quantification of mean Tnfa and Cxcr4b intensity (arbitrary units, arb. units) 50 μm proximal and distal to the wound border at 4 (n = 5 ventricles) and 10 (n = 7 ventricles) dpci. Data are presented as mean ± SD. P-values were calculated with unpaired two-sided t-tests. Source data are presented in the Source Data file. Gmrc1b in situ hybridization chain reaction (HCR) and immunostaining for GFP and F-actin in Tg(mpeg1:EGFP) ventricles at 10 dpci. The yellow dotted box denotes the zoomed image from the trabecular BZ and the blue dotted box denotes the zoomed image from the cortical BZ. *denotes background autofluorescence from erythrocytes. Scale bars: 100 μm in (A, C, E), and G), 20 μm in (Ai) and (Aii), (B), and zoomed images in (E and G).
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