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. 2011 Mar 17;286(20):18268–18276. doi: 10.1074/jbc.M110.201665

FIGURE 6.

FIGURE 6.

UVB-induced collagen cleavage inhibits HAS2 via αvβ3. A–D, human fibroblasts were grown in collagen gels to form dermal equivalents. A and B, time course of collagen neoepitope accumulation in response to UVB (10 mJ/cm2) as determined by immunostaining using collagen 2 3/4Cshort polyclonal rabbit antibody. Panel C indicates control. C, MMP1 mRNA expression. D, HAS2 mRNA expression. E and F, three-dimensional cultures of fibroblasts in collagen gels were subjected to irradiation with UVB (100 mJ/cm2). After 24 h, HAS2 mRNA was determined in the presence of the MMP inhibitor I (MMP 1 I., 300 nmol/liter) (E) or the αvβ3-blocking antibody LM609 (αvβ3 AB,5 μg/ml) or isotype control (Iso, 5 μg/ml) (F). G and H, polymeric collagen (120 μg/ml) was added to the medium of monolayer cultures of human skin fibroblasts (CS). Subsequently, the cultures were subjected to UVB irradiation (UVBS, 100 mJ/cm2) in the presence (G) or absence (H) of MMP inhibitor I (MMP 1 I.S, 300 nmol/liter). After 3 days, the conditioned medium was added to a new fibroblast culture in the presence or absence of MMP inhibitor I (300 nmol/liter) (G) or αvβ3-blocking antibody LM609 (αvβ3 ABS, 5 μg/ml) or isotype control (IsoS, 5 μg/ml) (H) for 24 h. Subsequently HAS2 mRNA was determined by real time PCR; n = 3, mean ± S.E., *, p < 0.05.