Fig. 2: Characterization of Retnla knockout in unsorted bone marrow cells exhibiting cardiac protective effect.

A The number of unsorted bone marrow cells (UBCs) was higher in Retnla knockout (KO) mice regardless of myocardial infarction (MI). B Cell survival and proliferation-related mediators were analyzed in UBCs isolated from WT and Retnla KO mice by western blot. C UBCs were isolated from WT and KO mice 2 days after MI, and mRNA levels of adiponectin (Adipoq), p21, and caveolin-1 (Cav1) were quantified by real-time PCR. D The number of phosphorylated histone H3 (pH3)-positive cells in bone marrow was quantified, and the representative images showed pH3-positive cells (red fluorescence) in the bone marrow. (WT MI 4d; n = 14, KO MI 4d; n = 11, WT MI 7d; n = 23, KO MI 7d; n = 23). Scale bar, 200 μm. E To compare the endogenous angiogenic potential of WT and KO mice, mesenchymal stem cells (MSCs) were used. MSCs were isolated from bone marrow and tube formation assay was performed. Tube length, tubular area, and the number of branching points were analyzed in MSCs from wild type and Retnla KO mice. Scale bar, 500 μm. F UBCs were isolated from WT (n = 12) or KO mice (n = 13), and injected into infarcted heart of wild-type mice (n = 12 in the WT to WT mice, n = 13 in the KO to WT mice). After 2 weeks, cardiac function was analyzed, and the representative echocardiograms were shown. G, H Immunofluorescence staining showed larger distributions of anti-inflammatory CD206(+) macrophages and vascular density in the KO mice than in the WT mice. EF ejection fraction, FS fractional shortening. Scale bars, 200 μm. Data are represented as mean ± SEM. #P < 0.05, ##P < 0.01, ###P < 0.001 (by Student’s t test or two-way ANOVA). The image of the mouse (http://clipart-library.com/clipart/724544.htm) is obtained from clipart library and used under the Creative Commons Attribution-Share Alike 4.0 International license (http://creativecommons.org/licenses/by-sa/4.0/).