TLR2/6-dependent stimulation by MALP-2 promotes angiogenesis in vivo. (A) Representative pictures of hematoxylin and eosin-stained sections of Matrigel plugs after the administration of MALP-2 (1 μg/mL) for 6 days. VEGF (50 ng/mL) was used as positive control. AB indicates antibody (25 μg/mL each). Insets represent the total Matrigel plug. Vascular structures are filled with erythrocytes. (B) Hemoglobin content is given in micrograms per milligram of Matrigel. (C) Vascular structures are expressed as percentage vascular area per field. Unspecific IgG (50 μg/mL) was used as antibody control (n = 4-8). **P < .01 vs control. #P < .05, ##P < .01 vs MALP-2. (B-C) Error bars represent mean ± SEM. (D) Immunofluorescent staining of Matrigel plaques after the administration of MALP-2 (1 μg/mL) for 6 days with antibodies against CD31, CD45, TLR2, and TLR6. Colocalization of CD31 with TLR2 and TLR6, respectively, was visualized by confocal microscopy (right panels). Representative pictures of 3 independent experiments are shown. Image acquisition: (A and left/middle panels of D) Leica DM 4000B microscope, 20×/0.50 NA dry objective, Leica DFC 320 camera, Leica QWin Version 3 software. (D right panels) Leica DM IRB microscope, 40×/1.25 NA oil objective, Leica immersion oil, TCS SP2 AOBS scanhead, Leica LCS confocal software, Version 1347.