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. 2011 Jan 10;192(1):171–188. doi: 10.1083/jcb.201008060

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© 2011 de Jesus Perez et al.

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Figure 4. — Production and secretion of FN is dependent on BMP-2–mediated activation of βC. (A and B) Quantitative RT-PCR (A) and Western immunoblotting (B) were used to measure changes in levels of FN mRNA and protein, respectively, in hPASMCs stimulated with 10 ng/ml BMP-2 over a period of 6 h. In B, levels of secreted FN were measured in hPASMCs transfected with either scrambled or two independent βC siRNAs. Densitometry values were expressed as the ratio of OD of FN relative to the Ponceau stain of an unrelated band in media samples concentrated by ultrafiltration. Bars represent means ± SEM from n = 3. **, P < 0.001; and ***, P < 0.0001 versus time 0 using one-way ANOVA with Dunnett’s. (C) Representative immunofluorescence images showing FN distribution in scrambled (top) or βC siRNA (bottom)–transfected hPASMCs stimulated with 10 ng/ml BMP-2 over 6 h. DAPI (blue) stain was used to label cell nuclei. Bar, 10 µm.