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. 2011 Nov 16;6(11):e27743. doi: 10.1371/journal.pone.0027743

Figure 7. Overexpression of Sox9 rescues calcification that occurs with Notch inhibition.

Figure 7

Porcine AVICs were cultured under four conditions: (A,E,I) untreated, (B,F,J) DAPT-treated, (C,G,K) Sox9 overexpression, and (D,H,L) DAPT-treatment following Sox9 overexpression. Bright field image of unstained cells (A-D), and cells stained with von Kossa (E-H) and Alizarin red (I-L) are shown. AVICs demonstrated calcification with calcific nodules in untreated and DAPT treated cells (A, B, E, F, I, J) but no calcification or nodules were found in Sox9 or DAPT/Sox9 treated cells (C, D, G, H, K, L). Three independent experiments were performed and representative images are shown. (M) Quantification of Alizarin red staining is shown. *, p value<0.05. (N) Schematic showing that Notch1-dependent activation of Sox9 is required for proper expression of valve ECM proteins that are essential for normal adult valve function. Conversely, loss of Notch1 or Sox9 results in dysregulated aortic valve ECM and CAVD.