Figure 3.

Increased expression of NBPF in affected SSc skin and in cultured SSc dermal fibroblasts. (A) Immunohistochemistry for NBPF on one representative skin biopsy from a healthy control (normal) or from a SSc patient. Note that fibroblasts within healthy control skin did not show any staining for NBPF (black arrows) whereas fibroblasts within SSc dermis were intensely stained in brown (arrow heads). Original magnification 20X. Panels are representative of five independent skin biopsies. Panels on the right are a tenfold magnification of the squared inset in the panels on the left. (B) Western blots of cell extracts from three different normal fibroblast cell lines and three different SSc dermal fibroblast cell lines cultured in monolayers with or without TGF-β1 (10 ng/ml) for 24 h employing an NBPF antibody. The NBPF bands shown have been identified based on their expected molecular weights. (C) Bar graph displaying a quantitative assessment of the data shown in (B). The gel blots used to prepare the images shown in (B) and (C) were cropped above the 130 KD and below the 70 KD molecular size marker bands because the migration of the relevant NBPF molecular species was expected to be between these molecular sizes. There were no other crops in the gel blots. Full blots of Western Blot included in this figure are shown in Supplementary Fig. S5.