Effect of α-solanine on the PANC-1 tumor xenograft growth and body weight of athymic nude mice.

<p>PANC-1 cells were subcutaneously injected into the flanks of nude mice. When the tumors were measurable, mice were given DMSO or α-solanine for 2 weeks. (A) Tumor volume/mouse as a function of time.(B) The mouse of each group was monitored for body weight once a day. Mean body weight/mouse as a function of time.(C) Tumor volume/mouse and (D) tumor weight/mouse at the end of the study were analyzed. *p<0.05, compared with the control.</p

In Vitro Angiogenesis Assay.

<p>(A) Representative micrographs (100×) of tube after treatment of conditioned medium for 6 h. HUVECS were plated onto the Matrigel –precoated wells and cultured in conditioned medium from PANC-1 cells treated with for 24 h. (B) The tube lengths were measured by Image-ProPlus 6.0. (C) The mRNA expression of VEGF was presented as means ± S.E.M. of three independent experiments.(D) Western blot analysis of the expression of VEGF. GAPDH was used as a loading control. (E) Quantification of the western blot result was performed by calculating the ratio of the value to the control group. *p<0.05, **p<0.01, compared with the control.</p

Additional file 1: of The rat pancreatic body tail as a source of a novel extracellular matrix scaffold for endocrine pancreas bioengineering

Figure S1. (A) Anatomical structure of the rat pancreas. (B) Schematic diagram of surgical procedures for rat pancreatectomy. (TIFF 2979 kb

Cell Viability Assay and Colony Assay.

<p>(A) Cells of each cancer cell line were treated with various concentrations of α-solanine for 24 h and 48 h. (B) Photos of anchorage-independent cell growth of PANC-1(100×magnification). (C) The colony number were quantified and data were calculated from three independent experiments. Each bar represents the means ± S.E.M. (n = 3). **p<0.01, compared with the control.</p

Prime pairs used in Realtime PCR.

<p>Prime pairs used in Realtime PCR.</p

Effect of α-solanine on phosphorylation of Akt and mTOR.

<p>PANC-1 cells were treated with various doses of α-solanine for 24 h, or 9 µg/µl of α-solanine for 6,12,18,24 h. (A,C) The phosphorylation level of Akt and mTOR were determined by Western blot. GAPDH was used as a loading control. (B,D) Quantification of the western blot results were performed by calculating the ratio of the value to the control group. *p<0.05, **p<0.01, compared with the control.</p

Effect of α-solanine on the expression of NF-κB/p65, β-catenin and TCF-1 in nucleus of PANC-1 cells.

<p>PANC-1 cells were treated with various doses of α-solanine for 24 h. (A,C) The level of NF-κB/p65, β-catenin and TCF-1 in nucleus were determined by Western blot. Laminb1 was used as a nuclear protein loading control. (B,D) Quantification of the densitometric results were performed by calculating the ratio of the value to the control group. *p<0.05, **p<0.01, compared with the control.</p

Effect of α-Solanine on expression of metastasis associated molecule.

<p>(A) The mRNA expression of MMP-2/9, EMMPRIN, CD44, ENOS and Ecadherin was presented as means ± S.E.M. of three independent experiments.(B) The expressions of MMP-2 and MMP-9 protein were analyzed by Western blot.(C) The expressions of Ecadherin protein was performed by Western blot.(D) Quantification of the western blot results were performed by calculating the ratio of the value to the control group.(E) The activity of MMP-2 and MMP-9 were analyzed by Gelatin Zymography. (F) Quantification of the Gelatin Zymography results were performed by calculating the ratio of the value to the control group. *p<0.05, **p<0.01, compared with the control. GAPDH was used as a loading control.</p

Additional file 2: of The rat pancreatic body tail as a source of a novel extracellular matrix scaffold for endocrine pancreas bioengineering

Figure S2. In vivo transplantation steps: (A-C) Procedure of recellularized pancreas transplantation. Firstly heparinization was accomplished through the injection of heparin into the inferior vena cava. Nephrectomy on the left kidney was performed with an empty kidney region, and the left renal artery and vein were retained as the grafted vessels. The recellularized pancreas was placed within the kidney region, and the arterial inlet was connected to a peristaltic pump to perfuse heparin sodium solution for several minutes before in vivo transplantation. The outlet of the recellularized scaffold was then connected to the recipient’s renal vein to perfuse the heparin sodium solution for several minutes to avoid thrombosis. The recipient’s renal vein and the inlet of the recellularized scaffold were blocked with vascular clips. The inlet of the recellularized scaffold was subsequently connected to the recipient’s renal artery. Finally, the vascular clips were removed, and whether the blood flow in the recellularized scaffold was unobstructed was observed. The incision was closed after confirmation that there was no bleeding around the transplanted pancreas. (TIFF 8673 kb

Cell Migration and Invasion Assays.

<p>Cells were treated with various concentrations of α-solanine for 24 h. (A) Cells were photographed(100×magnification). (B) The wound area were quantified in four fields in each treatment, and data were calculated from three independent experiments.(C) Cells were photographed(100×maginfication). (D) The invaded cells were quantified by counting fo DAPI-stained cells. Each bar represents the means ± S.E.M. (n = 3).*p<0.05, **p<0.01, compared with the control.</p