Figure 6

DUSP3 deficiency affects in vivo angiogenesis. (A-E) DUSP3^+/+ and DUSP3^−/− mice were injected subcutaneously in the flanks with 0.5 mL of Matrigel together with human b-FGF (250 ng/mL) and Heparin (0,0138 μg/mL). Ten days after injection, mice were sacrificed and Matrigels were removed. Representative photographs are shown in (A) from eight mice from each group. (B) Quantification of angiogenesis within the Matrigel plugs was achieved by measuring hemoglobin (Hb) concentration in the Matrigel homogenates and was reported as mg of measured Hb per mg of Matrigel. Results are presented as means ± SEM, n =15 in each group. (C) FITC-dextran was i.v. injected to DUSP3^+/+ and DUSP3^−/− mice 5 min before removal of subcutaneously implanted Matrigel plugs. Frozen sections were stained using Alexa 594 conjugated anti-CD31 and DAPI and visualized using fluorescent microscope. Representative micrographs of FITC-Dextran (green), CD31 (red), DAPI (blue) stainings and a merge of all are shown. (D-E) Quantification of CD31⁺ cells in blood vessel sections per mm² of Matrigel section (D) and FITC dextran intensity (arbitrary units) in Matrigel sections (E) from DUSP3^+/+ and DUSP3^−/− mice. Results are presented as means ± SEM, n = 10 in each group. Data are presented as mean ± SEM from 3 independent experiments (n = 15). (F-H) 10⁶ LLC cells were subcutaneously injected in the flank of DUSP3^+/+ and DUSP3^−/− female mice. 7 days later, tumors were removed and homogenized for Hb measurement. Representative photographs of the tumors are shown in (F) from six WT and seven DUSP3-KO mice. (G) Hb measurement in the tumors homogenates shown in F. (H) Weights of the tumors retrieved from the mice. Data are presented as mean ± SEM. *p < 0.05 (t-student test).