Figure 7
MFAP-4 over-expression prevents UVB-induced deterioration of collagen I.
(a) Immunohistochemical analysis with a rabbit anti-human collagen I antibody or a non-specific control rabbit IgG was carried out using paraffin embedded sections from the xenografted skin treated with a lentiviral vector encoding a control reporter gene with or without continuous UVB irradiation for 8 wks and using xenografted skin over-expressing MFAP-4 with UVB exposure for 8 wks. Scale bars = 100 μm. (b) Western blotting analysis with an anti-human collagen I or an anti-human β-actin antibody to assess the protein levels of collagen I in xenografted skin treated with lentiviral vectors encoding a control reporter gene or MFAP-4 before and after UVB exposure for 4 wks. (c) NHDFs were transfected with siRNAs for non-specific sequences (Control) or MFAP-4 twice during the culture for 8 days. Control siRNA-transfected cells were treated with or without 10 nM human recombinant MFAP-4 during the culture. Quantitative RT-PCR analysis of the MMP-1 mRNA expression in cultured NHDFs was performed as described in the legend of Fig. 5a. The values reported represent means ± SD. ***p<0.001. (d) Western blotting analysis with an anti-human MMP-1 antibody was conducted for supernatants from cultured NHDFs treated as described in Fig. 7c. The obtained bands of the active form of MMP-1 were analyzed using a densitometer and the values in the graph are represented relatively. The values reported represent means ±SD. **p<0.01, *p<0.05.
