Spermidine treatment within drinking water promotes autophagy through increased phosphorylation of ULK1 in mice independent of genotype but does not induce apoptosis. (A) Representative western blot of cerebellum from wild-type and MJD mice treated with vehicle or spermidine and probed for markers of autophagy p-ULK1, ULK, p62 and LC3B. (B) Quantification of the ratio between LC3II/I showed no significant differences after spermidine treatment. (C) Quantification of p62 showed no significant differences following spermidine treatment (D) Simple main effects analysis of p-ULK1 levels demonstrated a significant increase following spermidine treatment ( p = 0.0109, n = 6–7), independent of genotype. (E) Representative western blot of wild-type and MJD mice treated with vehicle or spermidine and probed for markers of apoptosis including cleaved- and total-caspase 3 and downstream substrates cleaved- and total-PARP1. (F) Quantification of cleaved- and total-caspase 3 showed no significant differences following spermidine treatment. (G) Quantification of the ratio between cleaved- and full-length-PARP1 revealed no significant differences following spermidine treatment. A two-way ANOVA was utilised for statistical analysis followed by Tukey post hoc analysis. Data represents mean ± SEM. * Represents p < 0.05
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