MHY2233 Attenuates Replicative Cellular Senescence in Human Endothelial Progenitor Cells via SIRT1 Signaling

Figure 1

Characterization of senescent EPCs: (a) passage 8 EPCs were considered young, and passage 18 EPCs were considered senescent. Young cells and senescent cells were cultured in medium, and SA-β-gal staining was performed. Representative images of SA-β-gal-stained young and senescent EPCs are shown. SA-β-gal-positive cells are stained a green color. Scale bars: 25 μM and 5 μM. (b) Quantification of SA-β-gal-positive cells from three replicates ( per group). (c) Young and senescent EPCs were seeded in 96-well plates, and cell proliferation capacity was determined using the WST-8 assay. Absorbance was measured at 0, 12, 24, and 48 h at 450 nM wavelength ( per group). (d) Young and senescent cells were seeded in the upper chamber of a Transwell® culture plate. After 6 h, migrated cells were stained and images were taken under a microscope at 100x magnification ( wells per group). (e) Quantification of the number of migrated cells among young and senescent EPCs using ImageJ software. (f) For tube formation capacity, young and senescent EPCs were seeded in 96-well plates containing Matrigel®. After 6 h, representative images were taken under a microscope at 40x magnification ( per group). Scale bars: 25 μM. (g) Quantification of the tube length of branches per field in μM and (h) number of tube branches per field using ImageJ software (). (i) Cells were harvested, and total cell lysates were subjected to western blot analysis for protein expression of SIRT1, acetylated p53, p16^INK4a, and p21 for young and senescent EPCs. , , and vs. young EPCs by unpaired Student’s -test.

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