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. 2015 Feb 26;64(7):2664–2675. doi: 10.2337/db14-1709

Figure 4.

Figure 4

Depletion of PLAC8 reduces proliferation and increases senescence. PLAC8-specific siRNA was used to deplete PLAC8 from GDM-exposed ECFCs, and functional assays were performed. Transfection of an siControl was used as the control. A: Western blotting confirms efficient PLAC8 protein knockdown. The Western blot shows that the control siRNA did not affect protein levels. Vinculin is the loading control. B and C: Cell cycle analysis was conducted using flow cytometry. B: Dot plots from a representative experiment are shown. C: Quantitation of cells in S phase shows increased proliferation with PLAC8 depletion (n = 12 using five different GDM-exposed ECFC samples; *P < 0.05 by paired t test). D: Representative image of siControl- and siPLAC8-transfected GDM ECFCs stained for senescence-associated β-galactosidase (blue cells). E: Quantitation of senescent cells demonstrates increased senescence with PLAC8 depletion (n = 5 experiments using two different GDM-exposed ECFC samples; *P < 0.05 by paired t test).