Cucurbitacin B Exerts Antiaging Effects in Yeast by Regulating Autophagy and Oxidative Stress

Figure 2

Effect of CuB on autophagy in yeast. (a) Effect of CuB on ATG2 and ATG32 gene expression after 12 h of treatment. (b, c) Effect of CuB on the replicative lifespan of K6001 and atg2 (b) and atg32 (c) mutants with K6001 background. (d) Fluorescent images of YOM38 yeast contained plasmid pR316-GFP-ATG8 and YOM36 yeast after treatment of RES or different doses of CuB in the SD medium and DAPI staining observed with a two-photon confocal fluorescent microscope. The upper line and middle line showed merged and DIC images of YOM38 yeast cells containing plasmid pR316-GFP-ATG8, respectively. The line below showed fluorescent images of YOM36. (e) Effect of CuB on the percentage of YOM38 cells containing plasmid pR316-GFP-ATG8 with free GFP. Three pictures containing more than 60 cells in each group were used for statistical analysis. (f) Western blot analysis of GFP-Atg8 and free GFP in yeast after shifting to the SD medium containing 0.3 μM CuB for different times. (g) Western blot analysis of GFP-Atg8 and free GFP in yeast after treatment with RES or CuB for 22 h in the SD medium. (h) Effect of CuB on the chronological lifespan of wild-type YOM36 and atg2 and atg32 mutants with YOM36 background. and indicate significant difference between treatment groups and the corresponding control group (, ).

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