Fig. 4

miR-200c is upregulated in DFU FBs and KCs and its targets are downregulated while p66Shc Ser-36 phosphorylation is decreased. Total RNA was extracted from FBs and KCs isolated from biopsies of DFU or biopsies of normoglycemic pts (HS). a miR-200c expression levels were induced in FBs isolated by biopsies of pts with DFU (FB DB) compared to HS (N = 7 FB HS, N = 8 FB DB; *P < 0.05). b miR-200c expression levels were induced in KCs isolated by biopsies of pts with DFU (KC DB) compared to HS (N = 8 KC HS, N = 10 KC DB; *P < 0.05). c ZEB1, SIRT1 and FOXO1 mRNA expression levels were downmodulated in FBs extracted by skin biopsies of pts with DFU vs healthy control subjects, while P66Shc mRNA was induced, albeit not significantly (N = 5 FB HS, N = 7–9 FB DB *p < 0.05; **p < 0.01). e ZEB1, SIRT1 and FOXO1mRNA expression levels were decreased in KCs isolated by skin biopsies of pts with DFU vs healthy control subjects, while P66Shc mRNA was induced (N = 5 KC HS, N = 7–8 KC DB *p < 0.05). e–f Representative western blots of ZEB1, SIRT1, FOXO1, P66Shc-P-Ser36 and P66Shc of DFU compared to HS ones of FBs and KCs, respectively. g–h Expression levels of ZEB1, SIRT1, FOXO1, P66Shc-P-Ser36 and P66Shc proteins were evaluated by densitometric analysis and normalized by either vinculin or α-tubulin protein levels, as indicated (N = 8 FB HS, N = 8 FB DB; N = 5–6 KC HS, N = 6 KC DB; *p < 0.05; **p < 0.01; ***p < 0.001).