The level of SHH expression was reduced in the cerebellums of Rere^om/eyes3 embryos at E18.5.

A. Protein extracts prepared from the cerebellums of Rere^om/eyes3 embryos and wild-type littermates at E18.5 were subjected to western blot analysis using an anti-SHH antibody and an anti-HSP70 antibody. Expression of SHH in the cerebellum of Rere^om/eyes3 embryos was lower than that of their wild-type littermates. Expression of HSP70 was comparable between Rere^om/eyes3 embryos and their wild-type littermates. B. To quantify the level of SHH expression, the density of the SHH band was normalized by the density of HSP 70 band in same blot. The expression level of SHH was significantly decreased in the cerebellums of Rere^om/eyes3 embryos when compared to the level of SHH in the cerebellums of their wild-type littermates at E18.5 (three independent experiments were performed for quantification; *  = p<0.01).