Fig. 6.

GSK-3β phosphorylation within a stented microenvironment in vitro and following vascular remodeling in vivo. a Representative immunocytochemical staining of pGSK-3β and Notch1 in bovine vSMC upstream and within the stented regions of a MVP following stent implantation after 4 days. Cells were visualised using a DAPI solution (4', 6-diamidino-2-phenylindole) and cell number determined by fluorescent microscopy in multiple random fields of view. Magnification 40× lm. b Cumulative data on the number of cells upstream and within the stented region of a MVP after 4 days in culture. Cells were counted blindly by determining the number of DAPI stained nuclei in situ from digital images of different constant regions of 300 × 300 pixels in random visual fields. Cells counts were confirmed using a hemocytometer following trypsinisation of individual cells. Data are the mean of four independent experiments performed in triplicate, *p < 0.05 vs upstream. c Representative immunohistochemical staining for pGSK-3β, total GSK-3β, α-actin, PCNA, Bax and Hrt-1 from ligated carotid arteries from C57B16/J mice after 14 days. d Quantitative realtime PCR analysis of GSK-3β mRNA levels from sham and ligated left carotid arteries from C57B16/J mice 3 and 14 days after ligation (4 vessels per preparation). RNA data were normalized to GAPDH mRNA levels. The cumulative data represents the mean values from three independent experiments ±SEM, *p < 0.05 versus sham control