1. Endothelial nitric oxide synthase affects both early and late collateral arterial adaptation and blood flow recovery after induction of hind limb ischemia in mice 
Brian Park, MD, Ari Hoffman, MD, Yagai Yang, PhD, Jinglian Yan, PhD, Guodong Tie, PhD, Hossein Bagshahi, MD, Philip T. Nowicki, MD, Louis M. Messina, MD 
Journal of Vascular Surgery 
Volume 51, Issue 1, Pages (January 2010)
DOI: /j.jvs
Copyright © 2010 Society for Vascular Surgery Terms and Conditions

2. Fig 1
Serum nitrite (NOx) concentration in wild-type, endothelial nitric oxide synthase (eNOS−/−) and early NG-nitro-l-arginine methyl ester (L-NAME) treatment groups. The serum NOx level was lower in the eNOS−/− mice than in the wild-type mice. L-NAME significantly reduced serum NOx levels ≤1 day after beginning L-NAME treatment, and the reduction was sustained throughout the duration of L-NAME treatment.
Journal of Vascular Surgery  , DOI: ( /j.jvs )
Copyright © 2010 Society for Vascular Surgery Terms and Conditions

3. Fig 2
Responses of wild-type, endothelial nitric oxide synthase (eNOS−/−) and early NG-nitro-l-arginine methyl ester (L-NAME) treatment groups to hind limb ischemia. A, Effects of femoral artery excision on hind limb tissue integrity are shown in photographs that are representative of all animals in each group. The ischemic hind limb is shown on the left and white arrows in eNOS−/− and L-NAME mice denote evidence of tissue necrosis. B, Laser Doppler perfusion imager (LDPI) data are shown. Each mouse underwent 3 scans at each time point, and the ratio of the LDPI signal from the ischemic and nonischemic hind limb was calculated; the average ratio was used as a single data point for each mouse. C, Collateral artery diameter on day 3 after the induction of hind limb ischemia was determined as the average of the largest arteries identified within 10 randomly selected low-power fields for each mouse by a blinded observer. The ratio of the average diameter in the ischemic thigh to the average diameter in the nonischemic thigh was determined for each mouse and used as a single data point.
Journal of Vascular Surgery  , DOI: ( /j.jvs )
Copyright © 2010 Society for Vascular Surgery Terms and Conditions

4. Fig 3
Presence of hemangiocytes in the thigh musculature in wild-type, endothelial nitric oxide synthase (eNOS−/−), and early NG-nitro-l-arginine methyl ester (L-NAME) treatment groups on day 3 after induction of hind limb ischemia. A, Quantitation of hemangiocytes, defined as cells that colocalized chemokine (C-X-C motif) receptor 4 (CXCR4) and vascular endothelial growth factor receptor (VEGFR) 1, was determined as the average number of hemangiocytes present in each mouse in 10 randomly selected high-power fields, determined by a blinded observer. B, In representative photomicrographs (original magnification ×200) of hemangiocyte immunostaining, VEGFR1 is stained green, CXCR4 is stained red, and nuclei are stained blue. A collateral vessel is present in each photomicrograph, evidenced by the linear cord of blue-stained nuclei. Note the presence of double-stained hemangiocytes clustered along the collateral artery in the ischemic thigh of the control mouse. C, Confocal microscopy of a collateral artery within the ischemic thigh of the wild-type group. The left photo is shown at original magnification ×200. The area delineated by the white square in the left photo is shown at original magnification ×400 in the right photo. Note the presence of a hemangiocyte in the adventitial area of a collateral artery. This adventitial distribution of hemangiocytes was commonly observed in the ischemic thigh muscle of wild-type mice, but it was never observed in eNOS−/− or early L-NAME treatment groups.
Journal of Vascular Surgery  , DOI: ( /j.jvs )
Copyright © 2010 Society for Vascular Surgery Terms and Conditions

5. Fig 4
Response of wild-type and late NG-nitro-l-arginine methyl ester (L-NAME) treatment groups to hind limb ischemia. A, Laser Doppler perfusion imager (LDPI) data. Each mouse underwent 3 scans at each time point, and the ratio of the LDPI signal from the ischemic and nonischemic hind limb was calculated; the average ratio was used as a single data point for each mouse. B, Collateral artery diameter on day 28 after induction of hind limb ischemia was determined as the average of the largest arteries identified in each mouse within 10 randomly selected low-power fields by a blinded observer. The ratio of the average diameter in the ischemic thigh to the average diameter in the nonischemic thigh was determined for each mouse and used as a single data point.
Journal of Vascular Surgery  , DOI: ( /j.jvs )
Copyright © 2010 Society for Vascular Surgery Terms and Conditions

6. Fig 5
Presence of hemangiocytes in the thigh musculature in control and late NG-nitro-l-arginine methyl ester (L-NAME) treatment groups 28 days after induction of hind limb ischemia. A, Quantitation of hemangiocytes, defined as cells that colocalized chemokine (C-X-C motif) receptor 4 (CXCR4) and vascular endothelial growth factor receptor (VEGFR) 1, was determined as the average number of hemangiocytes present in each mouse in 10 randomly selected high-power fields, determined by a blinded observer. B, In representative photomicrographs of hemangiocyte immunostaining, VEGFR1 is stained green, CXCR4 is stained red, and nuclei are stained blue. In contrast to the data obtained at 3 days after the induction of hind limb ischemia (Fig 2, B), note that hemangiocytes are present in the areas between myocytes; that is, at the sites of capillaries (original magnification ×200). C, Confocal microscopy of the ischemic thigh of the wild-type group. The left-hand photo is shown at original magnification ×200. The area delineated by the white square in the left photo is shown at original magnification ×400 in the right photo. Note the presence of hemangiocytes in the pericapillary region.
Journal of Vascular Surgery  , DOI: ( /j.jvs )
Copyright © 2010 Society for Vascular Surgery Terms and Conditions