d-PA, unlike n-PA, impairs EPC functions.

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Presentation transcript:

d-PA, unlike n-PA, impairs EPC functions. d-PA, unlike n-PA, impairs EPC functions. A: EPC proliferation in response to the indicated stimuli (data are expressed as mean + SD; n = 6; *P < 0.05, d-PA 48 h vs. C [control] 48 h; **P < 0.01, d-PA 72 h vs. C 72 h). B: FACS analysis indicating the percentage of EPCs at different cell-cycle phases (referred at 48 h) (data are expressed as mean, n = 6). C: FACS analysis of PCNA-positive EPCs treated as indicated (referred at 48 h). The reported graph is representative of six independent experiments. D: q-RT-PCR analysis of cyclin D1 and p21Waf mRNA expression in 48 h–treated EPCs as indicated. Data normalized to the corresponding glyceraldehyde-3-phosphate dehydrogenase mRNA and expressed as mean + SD are reported as fold induction relative to control values arbitrary set as 100 (n = 9; **P < 0.01, cyclin D1 d-PA vs. C and n-PA; ††P < 0.01, p21Wafd-PA vs. C and n-PA). E: Representative WB and densitometric analysis of cyclin D1 and p21Waf protein content in 48 h–treated EPCs. Densitometric data, normalized to the corresponding β-actin value and expressed as mean + SD, are reported as fold induction relative to control values arbitrarily set as 100 (n = 9; **P < 0.01, cyclin D1 d-PA vs. C and n-PA; †††P < 0.001, p21Wafd-PA vs. C and n-PA). IL-3– and AGE-stimulated ECs were used as positive controls for cyclin D1 or p21Waf, respectively. F: EPC migration in response to stromal cell–derived factor 1α (SDF-1α) (20 ng/mL). Data are expressed as mean + SD (n = 6; **P < 0.01, d-PA vs. C and n-PA). Antonella Trombetta et al. Diabetes 2013;62:1245-1257 ©2013 by American Diabetes Association