Research Paper Volume 13, Issue 11 pp 14687—14708

Age-related defects in autophagy alter the secretion of paracrine factors from bone marrow mononuclear cells

Heterochronic bone marrow reconstitution by Y-Sca-1+ BMCs is associated with enhanced autophagy in the aged mouse heart. (A) LC3I, II and p62 protein levels were measured in old mice hearts, reconstituted with GFP+young-Sca-1+ (Y+-O) or GFP+old-Sca-1+ (O+-O) bone marrow cells (BMCs) by Western blotting. (B, C) Band intensities shown in panel (A) were quantified to measure LC3II/I ratio or p62 levels. Data were normalized to loading control GAPDH. (D) Immunofluorescent staining was performed on sections of Y+-O and O+-O myocardium. LC3b is indicated by yellow, CD45 by red, GFP+ BMCs by green, and nuclei by blue. Scale bar represents 50 μm. Percentage of GFP+ cells (E), or LC3+ and GFP+ cells (F) in panel (D) were quantified. Scale bars represent 50 μm. T-test used to analyze data. n=5-6; *p≤0.05; **p≤0.01.

Figure 6. Heterochronic bone marrow reconstitution by Y-Sca-1+ BMCs is associated with enhanced autophagy in the aged mouse heart. (A) LC3I, II and p62 protein levels were measured in old mice hearts, reconstituted with GFP+young-Sca-1+ (Y+-O) or GFP+old-Sca-1+ (O+-O) bone marrow cells (BMCs) by Western blotting. (B, C) Band intensities shown in panel (A) were quantified to measure LC3II/I ratio or p62 levels. Data were normalized to loading control GAPDH. (D) Immunofluorescent staining was performed on sections of Y+-O and O+-O myocardium. LC3b is indicated by yellow, CD45 by red, GFP+ BMCs by green, and nuclei by blue. Scale bar represents 50 μm. Percentage of GFP+ cells (E), or LC3+ and GFP+ cells (F) in panel (D) were quantified. Scale bars represent 50 μm. T-test used to analyze data. n=5-6; *p≤0.05; **p≤0.01.