Lentiviral Vector–Induced Overexpression of NDNF Reduced Apoptosis of Old hBM-MSCs and Increased Angiogenesis
(A) Representative fluorescent micrographs show green fluorescent protein–positive (GFP+) cells (green) to indicate transduced cells. (B) NDNF mRNA expression was significantly higher in NDNF-transduced (NDNF+old) than in empty vector–transduced (old) hBM-MSCs, n = 6/group. The expression of NDNF protein (C) in cell lysates evaluated by Western blotting (n = 5/group) and (D) in the supernatant of cells evaluated by enzyme-linked immunosorbent assay (n = 6/group) was significantly higher in NDNF-transduced hBM-MSCs. (E) Representative immunofluorescent staining for terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL) (red) with nuclei stained blue with DAPI. Arrows indicate TUNEL+ cells. The percentage of TUNEL+ cells was significantly lower in NDNF-transduced cells than in empty vector–transduced cells, n = 3/group. (F) Western blot showed that the expression of the proapoptosis-related protein Bax was significantly lower in NDNF-transduced cells, n = 4/group. (G) Representative micrographs from the tube formation assay (red dots indicate tubular structure). The number of tubes formed was significantly higher in NDNF-transduced cells, n = 6/group. GAPDH = glyceraldehyde 3-phosphate dehydrogenase; other abbreviations as in Figure 1.