BUSCADOR RECOLECTA

(A) Western blot gel images of the three biological replicates used for the quantifications in Fig 5B and 5C, displaying Nup159-eGFP and free eGFP levels at the indicated time points after cells were transferred to SD-N medium (time = 0 h). Intermediate Nup159-eGFP degradation forms are indicated with (*). Pgk1 was used as a loading control. Graphs showing the quantification of the levels of full-length Nup159-eGFP and free eGFP for each of the experiments are also included next to each western blot image. Data are available in S1 Data., Peer reviewed

Supplementary Figure 1 – Principal coordinate analysis (PCoA) of the anterior-mid intestine microbiota similarity of gilthead seabream (Sparus aurata) fed experimental low fishmeal diets containing Debaryomyces hansenii (1.1%, 17.2 x 10+5 cfu; Yeast diet) or devoid of the probiotic (Control diet). -- Supplementary Table 1 – Summary of the differentially expressed genes (DEGs) obtained from the microarray-based transcriptomic analysis for the gut of juvenile gilthead seabream fed the D. hansenii diet. Table lists the DEGs descriptions and their corresponding acronyms, Fold Change (FC), gene regulation (up or down), and P-value. n/f defines the gene description for unknown genes. -- Supplementary Table 2 – Functional enrichment analysis of the DEGs. Biological process (Gene Ontology) identified in the intestine of juvenile gilthead seabream (Sparus aurata) fed the D. hansenii diet. -- Supplementary Table 3 – Top-10 biological process (Gene Ontology) of the DEGs identified in the intestine of juvenile gilthead seabream (Sparus aurata) fed the D. hansenii diet. -- Supplementary Table 4 – Representative cluster of the functional enrichment network for biological processes based on the related functions of the differentially expressed genes (DEGs) in the anterior-mid intestine of gilthead seabream (Sparus aurata) fed experimental low fishmeal diets containing Debaryomyces hansenii (1.1%, 17.2 x 105 cfu; Yeast diet) or devoid of the probiotic (Control diet). -- Supplementary Table 4 – Cellular component (GO) and annotated keywords (UniProt) of the DEGs identified in the intestine of juvenile gilthead seabream (Sparus aurata) fed the D. hansenii diet., Peer reviewed

(A–D) Stationary phase cultures in YPAD were diluted to OD600 = 0.2 in SD-N medium and grown for 24 h at 26°C. (A) Western blot gel images displaying Nup159-eGFP and free eGFP levels at the indicated time points are shown for otherwise wild-type, NUP159-eGFP and NUP159-eGFP bfa1Δ cells, all further carrying PEP4 and PRB1 gene deletions, after being transferred to SD-N medium (time = 0 h). Intermediate Nup159-eGFP degradation forms are indicated with (*). Pgk1 was used as a loading control. To facilitate visualization of the fainter bands overexposed images of the same gels are also shown (higher exposure). Experiment was carried out thrice (n = 3) and a representative image is shown. (B) Western blot gel images displaying Nup133-eGFP and free eGFP levels at the indicated time points are shown for wild type, NUP133-eGFP and NUP133-eGFP bfa1Δ cells after being transferred to SD-N medium (time = 0 h). Intermediate Nup133-eGFP degradation forms are indicated with (*). Pgk1 was used as a loading control. To facilitate visualization of the fainter bands overexposed images of the same gels are also shown (higher exposure). Experiment was carried out thrice (n = 3) and a representative image is shown. (C) Western blot gel images displaying Nup159-eGFP in wild-type cells, as well as levels of Nup159-AIM-eGFP in cells expressing Bfa1-GBP, in a bfa1Δ mutant or in an otherwise wild-type background, 24 h after being transferred to SD-N medium. Pgk1 was used as a loading control. Experiment was carried out thrice (n = 3) and a representative image is shown. (D) Quantification of the relative levels of free eGFP in (C). Data are the average of 5 experiments (n = 5) and are available in S1 Data. Error bars represent SEM. (E) ATG8 gene expression determined by quantitative RT-PCR in the indicated strains and normalized to the wild type. Data are the average of 3 experiments (n = 3) and are available in S1 Data. Error bars represent SEM. (F) Western blot gel images displaying GFP-Atg8 and free GFP levels 24 h after exponential cells were diluted in SD-N medium. Pgk1 was used as a loading control. Experiment was carried out 4 times (n = 4) and a representative image is shown. (G) Quantification of the relative levels of free GFP in (F). Data are the average of 4 experiments (n = 4) and are available in S1 Data. Error bars represent SEM., Peer reviewed

(A–F) Stationary phase cultures in YPAD were diluted to OD600 = 0.2 in SD-N medium and grown for 4 h (C–F) or 24 h (A, B) at 26°C. (A) Representative images of live cells expressing Nup192-eGFP (green) and Vph1-yomRuby2 (red) in atg15Δ and atg15Δ bfa1Δ cells. Phase-contrast (PhC) and merged images are also shown. (B) Quantification of the percentage of cells displaying (black bars) or not (white bars) extranuclear Nup192-eGFP foci. Data are the average of 3 experiments (n = 3; 100 cells/each) and are available in S1 Data. Error bars represent SD. (C–E) Quantification of the percentage of cells displaying extranuclear (C) and perinuclear (D) Nup159-eGFP foci, as well as of cells displaying an aberrant nuclear morphology (E), 4 h after being transferred to SD-N medium. An estimation of the percentage of cells displaying 1–2, 3–4, or more than 4 foci is also shown for both cells with extranuclear (C) and perinuclear (D) Nup159-eGFP clusters. Data are the average of 3 experiments (n = 3; 100 cells/each) and are available in S1 Data. Error bars represent SEM. (F) Quantification of the percentage of cells displaying (black bars) or not (white bars) extranuclear Nup159-eGFP foci. Data are the average of 3 experiments (n = 3; 100 cells/each) and are available in S1 Data. Error bars represent SD., Peer reviewed

Experimental chronogram: Figure S1: Chronogram of the glutathione manipulation. BSO = buthionine sulfoximine. From (Romero Haro and Alonso-Alvarez 2015). DNA extraction protocol. Supplementary statistical analyses. Supplementary Results: Table S1; Table S2; Table S3; Table S4; Table S5; Figure S2; Figure S3; Figure S4; Figure S5; Figure S6; Figure S7; Figure S8. Interaction between early glutathione levels and telomere length on longevity, Peer reviewed

List of the strains in this study, which also details the specific experiment in which each of the strains was used., Peer reviewed

Antibodies for immunofluorescence and western blot, Peer reviewed

List of primers used in this study for the analysis of gene expression by quantitative RT-PCR, Peer reviewed

Original images of all blots displayed in this study, Peer reviewed

Both the spindle microtubule-organizing centers and the nuclear pore complexes (NPCs) are convoluted structures where many signaling pathways converge to coordinate key events during cell division. Interestingly, despite their distinct molecular conformation and overall functions, these structures share common components and collaborate in the regulation of essential processes. We have established a new link between microtubule-organizing centers and nuclear pores in budding yeast by unveiling an interaction between the Bfa1/Bub2 complex, a mitotic exit inhibitor that localizes on the spindle pole bodies, and the Nup159 nucleoporin. Bfa1/Bub2 association with Nup159 is reduced in metaphase to not interfere with proper spindle positioning. However, their interaction is stimulated in anaphase and assists the Nup159-dependent autophagy pathway. The asymmetric localization of Bfa1/Bub2 during mitosis raises the possibility that its interaction with Nup159 could differentially promote Nup159-mediated autophagic processes, which might be relevant for the maintenance of the replicative lifespan., Peer reviewed