Supplemental Figure 1
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Fig. S1. Localization of RECK, type I collagen, and type II collagen at cartilaginous nodules formed by ATDC5 cells. ATDC5 cells incubated for 15 days were fixed and incubated in the absence (D,E) or presence of primary antibodies (A, anti-RECK; B, anti-Type I collagen; C, anti-Type II collagen) followed by rinsing and visualization with the secondary antibodies (A and D, TRITC-anti-mouse IgG; B,C,E, FITC-anti-rabbit IgG). Reconstituted z-axis images along two cutting lines (a,b) are shown in the bottom and right panels, respectively. Bars, 100 μm. Note that the fluorescent signals were minimal when primary antibodies were omitted (D,E) and that the single labeling images for RECK, type I collagen and type II collagen (A,B,C) support the conclusion based on Fig. 2E and 2F that type I collagen is enriched near the surface of the nodules whereas RECK and type II collagen are abundant inside the nodules.
