Quantitative fluorescent Western blots validate changes in cell cycle proteins in Wldscerebellum in vivo. Bar chart showing percentage change in protein expression (mean ± standard error of the mean; n ≥ 3 for all proteins) in Wldscerebellum compared with wild-type. As expected, Wlds protein expression was highly upregulated (left bar). The second portion of the graph shows increases in both pituitary tumor transforming gene 1 (Pttg1) and ubiquitin-activating enzyme E1 (Ube1) proteins in Wldsmice, both of which have previously been implicated in the Wldsneuroprotective phenotype [22, 31]. The third portion of the graph shows validation for two genes highlighted on the SuperArray analysis as being upregulated by more than twofold. The final portion of the graph shows similar increases in cell cycle proteins not included on the SuperArray plate, showing that increased expression of cell cycle proteins is not restricted to those included on the SuperArray. Statistical tests were carried out comparing raw expression data from wild-type mice with those from Wldsmice. **P < 0.01, P < 0.001 by unpaired t-test (two-tailed). ns, not significant.