Fig 1: BX795 restores neurodegenerative phenotypes via the mTORC1 signaling pathway.a Representative images of SH-SY5Y cells stably transduced to express DsRed only or DsRed and human pA53T-αSyn. After neuronal differentiation, cells were immunolabled for αSyn (SNCA), TUJ1 and pRPS6. b Western blot showing that the presence of mutant SNCA in differentiated p.A53T-transduced SH-SY5Y cells, results in an increase in the levels p-mTOR and p-RPS6. Actin shows equal protein loading. Data represent mean ± SEM (t test, *P < 0.05, n = 3 independent experiments). c Western blot showing an acute reduction in the levels of p-mTOR and p-RPS6 in the above stably transduced and differentiated SH-SY5Y cells, in the presence of BX795. Actin shows equal protein loading. Data represent mean ± SEM (Comparisons by ANOVA with Tukey correction, *P < 0.05, ****P < 0.0001, n = 3 independent experiments). d BX795 inhibits the kinase activity of pure recombinant p70 S6K (25 ng/assay) in a dosedependent manner. Rapamycin (Rapa) was used as a positive control. Data represent mean ± SEM (Comparisons by ANOVA with Tukey correction, *P < 0.05, **P < 0.01, n = 3). e Western blot showing the effect of BX795 treatment alone and in combination with mTOR activator MHY-1485 on the p(Ser129)-αSyn levels in SH-SY5Y cells stably transduced to express DsRed only or DsRed and human pA53T-αSyn. f Representative confocal images show protein aggregates in control (ctl) and patient p.A53T neurons, either non-treated (DMSO) or treated with BX795 alone or in combination with the mTOR activator MHY-1485.