Figure 6: MT1-MMP restrains VEGF-C production in macrophages.

(a) Real-time qPCR analyses of Vegf-c transcripts in BMMs from WT and Mmp14^−/− mice on LPS (1 μg ml⁻¹) stimulation (**P<0.01, ***P<0.001, n=3, two-tailed t-test). Western blotting analyses of VEGF-C protein expression (b) and real-time qPCR analyses of Vegf-c mRNA expression (c) in thioglycollate-elicited macrophages in WT mice transplanted with either WT or Mmp14^−/− bone marrow. Quantification of VEGF-C protein expression was shown in the right panel of b (*P<0.05, ***P<0.001, n=3, two-tailed t-test). (d) Morphometric comparison of corneal lymph vascularized areas in WT mice transplanted with either WT or Mmp14^−/− bone marrows (**P<0.01, n=5, two-tailed t-test). Scale bars, 400 μm. (e) qPCR analyses of Vegf-c mRNA levels in corneas from WT mice transplanted with either WT or Mmp14^−/− bone marrows (**P<0.01, n=5, two-tailed t-test). (f) Western blotting analyses of MT1-MMP in Mmp14^flox/floxLysM-cre(−) and Mmp14^flox/floxLysM-cre (+) peritoneal macrophages (n=4). (g) Morphometric comparison of corneal lymphvascularized areas in Mmp14^flox/floxLysM-cre (−) and Mmp14^flox/floxLysM-cre (+) mice at P15. The quantification of vascularization area were shown in h (*P<0.05, n=5, two-tailed t-test). Scale bars, 400 μm. (i) Real-time qPCR analyses of Vegf-c mRNA levels in corneas from Mmp14^flox/floxLysM-cre (−) and Mmp14^flox/flox LysM-cre (+) mice at P15 (**P<0.01, n=3, two-tailed t-test). (j) Whole-mounted corneas from P15 Mmp14^flox/floxLysM-cre (−) and Mmp14^flox/flox LysM-cre (+) mice were immunostained using VEGFR-3 (red) and CD11b (green) antibodies (n=5). Scale bars, 25 μm Data represent the mean±s.e.m. The experiments were repeated at least three times.