Fig 5. Autophagy inhibition enhanced α-syn-associated toxicity in strain A3.

(A) The A1-A4 strains, which lack the gene encoding Atg5, were monitored for α-syn mediated toxicity as described above in Fig 1. In contrast to wt, the corresponding strains of A3 and A4 lacking Atg5 grew poorly upon α-syn overexpression. (B) The yeast strains transformed with plasmid encoding α-syn under a galactose inducible promoter were grown in inducing media. At indicated time, cells were collected, lysed, and the lysate probed with anti α-syn antibodies. The α-syn abundance was found to be lower in strain A3 than in strain A2 at all time points. (C) The indicated strains transformed with plasmid encoding α-syn under a galactose inducible promoter. Cells were grown under inducible conditions for 12 h. The qRT-PCR was carried out with primers specific for the genes encoding α-syn or Pgk1 (internal control). (D) The steady state level of α-syn was measured in whole-cell lysates obtained from the indicated strains. As seen, α-syn levels were up-regulated upon deletion of the Atg5 encoding gene (atg5Δ/A3). Error bars represent the standard error of replicates performed 3 times. P-values were calculated using paired t-test.