Asporin Reduces Adult Aortic Valve Interstitial Cell Mineralization Induced by Osteogenic Media and Wnt Signaling Manipulation In Vitro

Figure 3

Endogenous Asporin mRNA and protein expressions decrease upon osteogenic induction in avian adult aortic valve interstitial cells (AVICs) in vitro. (a) (I and II): reduced Asporin gene expression in osteogenic (OS) media-induced AVICs compared to uninduced control AVICs, by reverse-transcriptase PCR and real-time PCR. Here, Asporin has shown reduced expression in OS-treated AVICs compared to control AVICs (a, I). Also, quantitative expression data reveals 0.62 fold decreased expression of Asporin in OS-treated AVICs compared to control AVICs (a, II). (b) (I and II): reduced Asporin protein expression in osteogenic (OS) media-induced AVICs compared to uninduced control AVICs, by western blot analyses. Here, Asporin has shown decreased expression in OS-treated AVICs compared to control AVICs, according to band intensity (b, I). Also, quantitative expression data reveals 0.67 fold decreased expression of Asporin in OS-treated AVICs compared to control AVICs (). (c) (I and II): here, AVICs were immunostained with Asporin antibody (green) and nuclei were counterstained with DAPI (blue), showing decreased number of Asp+ cells in OS-treated AVICs compared to control AVICs (marked by arrowheads). (d) Here, quantitative data demonstrate that OS-treated AVICs show decreased number of Asporin-positive cells as detected by 12.65% compared to untreated control AVICs. Statistical significance was determined by Student’s -test, where denotes and denotes and .

(a)

(b)

(c)

(d)