Research Paper Volume 10, Issue 9 pp 2407—2427

Heat-induced longevity in budding yeast requires respiratory metabolism and glutathione recycling

Figure 4. Yap1p is required for the mild heat shock induced RLS extension. (A) Oxygen consumption was increased under heat shock in the YAP1Δ strain. Oxygen uptake was measured polarographically using a Clark-type electrode equipped oxygraph. (B) Mitochondrially produced superoxide (measured by FACS as MitoSOX fluorescence in 10000 cells) was increased in the YAP1Δ strain during heat shock. The results are presented as the percentage of MitoSOX fluorescence detected in WT strain at optimal growth temperature. (C) qPCR measurement of gene expression levels showed that the absence of YAP1 did not affect the heat shock induced metabolic changes observed in WT. (D) NADP+/NADPH ratio was decreased during heat shock in the YAP1Δ strain. (E) GSH levels were not affected by heat shock in the YAP1Δ strain. (F) Mean and maximum replicative lifespan (RLS) were decreased in YAP1Δ already at OGT, and further decreased following heat shock. The number of curated cells is 74 for YAP1Δ and 84 for YAP1Δ HS. Unless otherwise stated, data in graphs are mean ± SEM from three biological and three technical replicates. ***P < 0.001; **P < 0.01; *P < 0.05 (ANOVA plus post hoc).