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. 2000 Sep 4;150(5):1085–1100. doi: 10.1083/jcb.150.5.1085

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© 2000 The Rockefeller University Press

This article is distributed under the terms of an Attribution–Noncommercial–Share Alike–No Mirror Sites license for the first six months after the publication date (see http://www.rupress.org/terms). After six months it is available under a Creative Commons License (Attribution–Noncommercial–Share Alike 4.0 Unported license, as described at http://creativecommons.org/licenses/by-nc-sa/4.0/).

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The effect of rhBMP-2 on the expression of desmin and alkaline phosphatase by the mc13 cells. Desmin staining of freshly isolated mc13 clones (A) showed that all of these cells express desmin. The phase-contrast view (B) is shown to demonstrate that a high percentage of the cells were desmin positive. Mc13 cells were incubated in growth media containing 200 ng/ml rhBMP-2 for 6 d. Cells were then stained for desmin expression, and percent desmin positive cells calculated by visualization with immunofluorescence. As a control, mc13 cells were grown in parallel without addition of rhBMP-2. When grown without rhBMP-2, mc13 cells remained uniformly (90–100%) desmin positive (C). With exposure to rhBMP-2, there is a significant decrease (*P < 0.05) in relative number of desmin positive cells (30–40%) within 6 d (C). The npmc and mc13 cells were also analyzed for expression of alkaline phosphatase after 6 d of growth with and without exposure to 200 ng/ml rhBMP-2. With rhBMP-2 stimulation, mc13 cells show a >600-fold increase in alkaline phosphatase activity than nonstimulated cells (D). The npmc show only minimal alkaline phosphatase activity with or without rhBMP-2 (D). *, Indicates a significant difference using the t test (P < 0.05). Bar, 100 μm.

The effect of rhBMP-2 on the expression of desmin and alkaline phosphatase by the mc13 cells. Desmin staining of freshly isolated mc13 clones (A) showed that all of these cells express desmin. The phase-contrast view (B) is shown to demonstrate that a high percentage of the cells were desmin positive. Mc13 cells were incubated in growth media containing 200 ng/ml rhBMP-2 for 6 d. Cells were then stained for desmin expression, and percent desmin positive cells calculated by visualization with immunofluorescence. As a control, mc13 cells were grown in parallel without addition of rhBMP-2. When grown without rhBMP-2, mc13 cells remained uniformly (90–100%) desmin positive (C). With exposure to rhBMP-2, there is a significant decrease (*P < 0.05) in relative number of desmin positive cells (30–40%) within 6 d (C). The npmc and mc13 cells were also analyzed for expression of alkaline phosphatase after 6 d of growth with and without exposure to 200 ng/ml rhBMP-2. With rhBMP-2 stimulation, mc13 cells show a >600-fold increase in alkaline phosphatase activity than nonstimulated cells (D). The npmc show only minimal alkaline phosphatase activity with or without rhBMP-2 (D). *, Indicates a significant difference using the t test (P < 0.05). Bar, 100 μm.