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. 2015 Nov 26;5:16872. doi: 10.1038/srep16872

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Copyright © 2015, Macmillan Publishers Limited

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(a) Fibulin-4^+/R and Fibulin-4^R/R SMCs show equal Tgf-β1 mRNA expression levels compared to Fibulin-4^+/+ SMCs. (b) Increased TGF-β1 levels measured in conditioned medium (CM) from Fibulin-4^R/R SMCs compared to Fibulin-4^+/+ CM on day 1–4 after serum starvation. Fibulin-4^+/R SMCs showed significant increased TGF-β1 levels on day 2 and 4 after serum starvation compared to Fibulin-4^+/+ SMCs. Furthermore, on day 4 Fibulin-4^R/R SMCs show significant increased TGF-β1 levels compared to Fibulin-4^+/RSMCs (n = 4 per day for each genotype). Two-way ANOVA analysis for genotype and between days p < 0.05. (c) Gradually increased TGF-β1 is also observed in aortic arch lysates of Fibulin-4^+/R (n = 6) and Fibulin-4^R/R mice (n = 5) compared to Fibulin-4^+/+ aortas (n = 5). This increase is significant in Fibulin-4^R/R aortic arch lysates compared to Fibulin-4^+/+ aortic arch lysates. (d) Fibulin-4^+/R and Fibulin-4^R/R SMCs show gradual increased Tgf-β2 mRNA expression levels compared to Fibulin-4^+/+ SMCs. (e) Measurement of TGF-β2 revealed markedly increased levels in CM of Fibulin-4^+/R and Fibulin-4^R/R SMCs, while TGF-β2 was undetectable in CM of Fibulin-4^+/+ SMCs (experiments were performed in at least 3 independent experiments). Two-way ANOVA analysis for genotype and between days p < 0.05. (f) Measurements in aortic arch lysates display significantly increased TGF-β2 in Fibulin-4^R/R aortas compared to Fibulin-4^+/R and Fibulin-4^+/+ aortas (*p < 0.05, **p < 0.01).