FIG. 3.

D-type cyclin protein regulation in Shh-treated versus vehicle-treated cultures. (A) Quantification of changes in cyclin D1 protein levels in the presence or absence of Shh for the indicated intervals. Western blots were quantified by densitometry, and values are expressed as the fold differences. The asterisks indicate significant differences between Shh- and vehicle-treated CGNPs at P < 0.01 (∗∗) and P < 0.05 (∗) confidence levels. (Inset, left) Immunohistochemical analysis of Shh-treated CGNPs showing overlap (yellow) of antibodies against cyclin D1 (Cy-2, green) and Math-1 (Cy-3, red). All cyclin D1-positive cells were also labeled with Math-1. The adjacent panel (inset, right) shows DAPI staining of the corresponding field of cells. (B to D) Representative time course of D-type cyclin protein level regulation by Shh. (B) Cyclin D1 appeared as a doublet with a relative mobility of 32 to 34 kDa. (C) Compared with controls, cyclin D2 levels did not appear to increase until after approximately 24 h. (D) Cyclin D3 levels were unaffected under the culture conditions tested.