Additional file 1: Table S1. of Attenuation of teratoma formation by p27 overexpression in induced pluripotent stem cells

Specific primer sequences for RT-PCR analyses. (DOCX 12 kb

Additional file 3: Figure S2. of Attenuation of teratoma formation by p27 overexpression in induced pluripotent stem cells

Effects of p27 overexpression on EB formation. Percentages of wells which have EBs and beating EBs are shown in A and B, respectively. Error bars correspond to the SEM (n = 3). *P < 0.05, student’s t-test. (PDF 341 kb

Additional file 4: Figure S3. of Attenuation of teratoma formation by p27 overexpression in induced pluripotent stem cells

Expression profile of stem cell factors in miPSCs-p27. RT-PCR analysis of stem cell marker genes (A) and relative intensities (B) of miPSCs and miPSCs-p27 are shown. Error bars correspond to the SEM (n = 3). *P < 0.05, student’s t-test. (PDF 311 kb

Additional file 2: Figure S1. of Attenuation of teratoma formation by p27 overexpression in induced pluripotent stem cells

Purification of p27 expressing miPSCs. Flow cytometric analysis of miPSCs (A), or p27-2A-mRFP stable transfected miPSCs (miPSCs-p27) before (B) and after (C) purification by cell sorter. Intensities of mRFP fluorescent were plotted against side scatters (SSC). The area surrounded by red line in B was sorted. (PDF 348 kb

Changes in ROS production.

<p>(A) PMA stimulated ROS production in PMNCs. Neutrophils were stimulated by 10 nmol/L PMA and ROS production was measured by Flow-cytometer as the intensity of DCFH fluorescence of 3000 cells. Bar graphs represent mean fluorescent intensity ± SD (*<i>p</i><0.05 vs control with PMA stimulation, †<i>p</i><0.05 vs AII with PMA stimulation, n = 6–8 for each group). (B–D) Superoxide production in the aortic wall. In situ detection of vascular superoxide production in Aorta (B), femoral arteries without cuff (C) and with cuff injury (D). Tissue superoxide was detected with DHE. Photomicrographs show representative results from 4 separate experiments. Scale bars indicate 50 µm in (B) and 500 µm in (C, D). Bar graphs represent mean fluorescent intensity ± SD (*<i>p</i><0.05 vs Control, †<i>p</i><0.05 vs BSO and AII, n = 7 of each). All statistical comparisons were evaluated using one-way ANOVA followed by Bonferroni post hoc analysis.</p

BSO inhibited AII induced aorta medial thickness.

<p>(A) Representative micrographs of cross sections of aorta 4 weeks after treatment. Hematoxylin and eosin staining: magnification ×400, Scale bars indicate 500 µm. (B) Graphs shows medial thickness of Aorta. Bar graphs represent mean ± SD (*<i>p</i><0.05 vs control, n = 7 of each).</p

BSO inhibited AII induced medial thickness of coronary artery.

<p>(A) The effects of AII and BSO on coronary arteries. Representative micrographs of cross sections of coronary arteries with hematoxylin and eosin staining. : magnification ×400, Bar indicates 100 µm. (B) Graphs shows coronary wall to lumen ratio. Bar graphs represent mean ± SD (*<i>p</i><0.01 vs AII+BSO, n = 7 of each).</p

Circumferential Vascular Wall Stresses.

<p>Graphs shows mean circumferential vascular wall stress (σ_cMAP) (A) and systolic circumferential vascular wall stress (σ_cSAP) (B). Values were calculated using Laplace theorem from blood pressure, lumen radius and wall thickness. Bar graph represent mean ± SD (*<i>p</i><0.01 vs BSO, †<i>p</i><0.01 vs AII, n = 7 of each).</p

Localization of gp91-phox in cuff-injured arteries.

<p>(A) Representative immunohistochemical photomicrographs of femoral artery cross sections. 1 weeks after cuff injury, arteries were stained with anti-gp91 phox antibody: cuff injured control artery (Control), treated with BSO (BSO), AII (AII), and AII and BSO (AII+BSO). Reactions were stained with horseradish peroxidase substrates for color development, with diaminobenzidine in brown. Scale bars indicate 200 µm. (B) Graph shows ratio of apoptotic cells in intima at 4 weeks after cuff injury. Bar graphs represent mean ± SD (*<i>p</i><0.01 vs AII+BSO, n = 5 of each).</p

Body weight, Heart rate, Left Ventricular Weight/Body Weight Ratio, Whole Blood GSH/GSSG Concentration, Plasma GSH Concentration.

<p>Body weight, Heart rate, Left Ventricular Weight/Body Weight Ratio, Whole Blood GSH/GSSG Concentration, Plasma GSH Concentration.</p