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. 2017 Mar 3;7:46. doi: 10.1038/s41598-017-00163-2

Figure 2.

Figure 2

Effects of ucOCN on tunicamycin-induced ER stress and impaired insulin signaling in HUVECs. Tunicamycin (Tun) was used to induce insulin resistance. To determine the effects of uncarboxylated osteocalcin, HUVECs were treated with 5 ng/ml of uncarboxylated osteocalcin for 4 h. For insulin signaling, cells were stimulated with 10 nM of insulin for 10 min. The relative quantity of proteins was analyzed using Quantity One software. (A) Phosphorylation of PERK and eIF2α in HUVECs. (B) Densitometric analyses of PERK and eIF2α in HUVECs. (C) IRS-1 tyrosine phosphorylation and Akt Ser-473 phosphorylation in HUVECs. (D) Densitometric analyses of IRS-1 tyrosine phosphorylation and Akt Ser-473 phosphorylation in HUVECs. A representative blot from three independent experiments is shown and the data expressed as mean ± SEM in each bar graph represent the average of three independent experiments. *P < 0.05 (Tun vs. control). #P < 0.05 (Tun/ucOcn vs. Tun). IB, immunoblot; IP, immunoprecipitation.