Fig. 1.

A PMSCs in high-density monolayer were stained for autophagy-associated vacuoles and DAPI. The %-positive area of autophagy stain was normalized to DAPI to quantify the relative change of autophagy level. At day 4, the PMSCs with 10nM rapamycin showed a marked increase in autophagy. B The rapamycin treated cells had a 3.94-fold increase in autophagy-associated stain compared to the vehicle control (p = 0.004, n = 5). C High-density PMSC monolayers were stained for senescence-associated β-galactosidase activity on day 4 and 7. D At day 4, the rapamycin group had 67.7% less SA β-Gal positive area (p = 0.0067, n = 15). At day 7, the rapamycin group had 72.9% less SA β-Gal positive area (p = 2.16x10⁻¹¹, n = 15). E Forward scatter and (F) side scatter were used as measures of relative cell size and complexity. The rapamycin group had a 5.52% and 8% decrease in forward scatter at day 4 and 7, respectively. Additionally, side scatter decreased by 3.4% and 4.7% at day 4 and 7, respectively. Note: *p < 0.05, **p < 0.01, ***p < 0.0001