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. 2019 Sep 13;8:e48240. doi: 10.7554/eLife.48240

Figure 5. Enforced expression of Cln3 increases lifespan in a chaperone-dependent manner.

(A) Survival curves of control and CLN3 overexpressing cells (n > 300). Curves predicted by the integrative model in Figure 4A are also shown (inset). (B) Budding volume distributions (n > 250) of young or old cells pre-aged for 24 hr before induction of CLN3 expression. (C) Lifespan effects of CLN3 overexpression in young (n = 200) or old cells pre-aged for 24 hr (n = 150) and 48 hr (n = 100). Median ±Q values are also plotted. (D) Survival curves of wild-type, Ydj1-deficient and SSA1 YDJ1 overexpressing cells (n > 300). Curves predicted by the integrative model in Figure 4A are also shown (inset). (E) Simulation of free Ydj1 and ProtM aggregate numbers during successive replicative cycles in wild-type cells by the integrative model. The results of a representative run in stochastic mode are shown. A 2-order of magnitude increase in Ydj1 levels (blue arrow) would only cause a very small delay (orange arrow) in protein aggregation and, as a consequence, in lifespan. (F) Lifespan of wild-type (wt) and Ydj1-deficient cells with empty vector (ydj1) or overexpressing CLN3 (ydj1 oCLN3) (n = 150). Median ±Q values are also plotted. The plot on the right shows the predicted lifespan obtained from independent simulations (n = 75) for the corresponding genotypes. Shown p-values were obtained using a Mann-Whitney U test. Results shown in this figure are representative of at least two replicate experiments.

Figure 5—source data 1. Enforced expression of Cln3 increases lifespan in a chaperone-dependent manner.
DOI: 10.7554/eLife.48240.021

Figure 5.

Figure 5—figure supplement 1. Lifespan analysis by MEP-induced microcolony size.

Figure 5—figure supplement 1.

(A) Schematic of a microcolony formed by the accumulation of G2-arrested daughter cells (orange) produced after MEP induction by a single mother cell (M, green) during successive division cycles. (B) A representative image (left) of microcolonies of different size formed under MEP-inducing conditions. ImageJ-delimited microcolony areas used to estimate the number of daughter cells produced and, hence, the number of generations, are also shown (right). (C) Microcolony size (mean ±CL) as a function of pre-aging time before plating. (D) Survival curves of wild-type and cln3 mutant cells estimated from MEP-induced microcolony sizes (n > 250). Bar = 20 µm.