BUSCADOR RECOLECTA

able S1: Species, sample, coverage, non-conversion rate (%), and average DNA methylation levels (mCG/CG) for all datasets generated in this study. Table S2: Genomic location of all differentially methylated regions (DMRs) identified in the medaka genome. Table S3: DMR-linked genes in zebrafish and medaka genomes., Peer reviewed

40 pages, Supplementary information for the article https://doi.org/10.1038/s41598-021-02328-6, Approach.-- Standardization of indicators,-- Calculation of index values.-- Socioeconomic indicators and factors.-- Institutional Indicators and Factors.-- Resilience factors summary.-- Index performance.-- Additional analyses.-- Bibliography, Peer reviewed

Figure S1. Identification of flagellaless GR4flaAB-derivative transposants impaired in the response to volatile 2-tridecanone (2-TDC). Figure S2. Multiple sequence alignment by MUSCLE of DnaJ amino acid sequences from different bacterial species Figure S3. Growth of S. meliloti dnaJ mutants and their parental strains on solid and liquid media. Figure S4. Effect of H2O2 on S. meliloti GR4 and GR4flaAB cell survival. Figure S5. Appearance of alfalfa plants inoculated with S. meliloti dnaJ mutant strains at the end of the nodulation kinetics experiment. Figure S6. Complementation of the symbiotic phenotype of S. meliloti dnaJ mutants. Table S1. Bacterial strains and plasmids used in this study. Table S2. List of primers used in this study, Peer reviewed

Supplementary Figure 1. GFP+ cells engraftment and mFVIII production correlation in transplanted mice. Supplementary Figure 2. Bleeding assay in newborn HA following transplantation. Supplementary Figure 3. Bleeding assay in adult HA following transplantation. Supplementary table 1. List of antibodies used for immunofluorescence and flow cytometry. Supplementary table 2. Engraftment (% GFP+ cells) and correction (% mFVIII activity) levels in plasma of newborn mice according to BU dosage and cell transplantation., Peer reviewed

Despite the great advances in macroautophagy/autophagy research in the last years, the in vivo role of the different members of the four mammalian orthologs of yeast Atg4 protease (ATG4A-D) remain unclear. To gain further insights into the functional relevance of Atg4 orthologs, we have generated mutant mice deficient in Atg4c. These mice are viable and fertile, and do not display any obvious abnormalities, indicating that they are able to develop the autophagic response required during the early neonatal period. However, they show tissue-specific autophagy alterations, including reduced autophagic flux in diaphragm and show decreased breathing and locomotor activity after fasting. In addition, atg4c-/- mice show reduced number of circulating T and B lymphocytes, which is associated with accumulation of apoptotic cells in the spleen and an increased susceptibility to develop chemically-induced fibrosarcomas. Moreover, through the analysis of cells and mice simultaneously deficient for ATG4C and ATG4D proteases we also reveal a role for ATG4C in mATG8 proteins delipidation. ATG4 (autophagy related 4 cysteine peptidase); ATG4A (autophagy related 4A cysteine peptidase); ATG4B (autophagy related 4B cysteine peptidase); ATG4C (autophagy related 4C cysteine peptidase); ATG4D (autophagy related 4D cysteine peptidase); Atg8 (autophagy related 8); GABARAP (GABA type A receptor-associated protein); GABARAPL1(GABA type A receptor-associated protein like 1); GABARAPL2 (GABA type A receptor-associated protein like 2); MAP1LC3A/LC3A (microtubule associated protein 1 light chain 3 alpha); MAP1LC3B/LC3B (microtubule associated protein 1 light chain 3 beta); mATG8 (mammalian Atg8); PE (phosphatidylethanolamine); PS (phosphatydylserine); SQSTM1/p62 (sequestosome 1)., This work was supported by grants from Ministerio Ciencia eInnovación (Spain) (PID2021-127534OB-I00), the South-Eastern 1315 Norway Regional Health Authority (2021088 to N.E.) and Instituto de Salud Carlos III (RTICC Spain). Jesús Prieto-Lloret is funded by Programa Estrategico IBGM, Escalera de Excelencia, ref. CCVC8485, Consejería de Educación, Junta de Castilla y León (Spain). Funding for open Access Charge: Roche Farma”, as the aricle will be published via Open access and the OA costs will be funded by Roche Farma., Peer reviewed

(A) An asynchronous culture of GAL-SIC1ΔNT GLN3-9MYC (YMF4471) was grown at 30 °C in medium lacking galactose. After the addition of nocodazole, culture was synchronised in G2-M phase for 1 generation time. Cells were then transferred to fresh medium containing galactose to allow overexpression of SIC1ΔNT. Cells were maintained as well as in nocodazole. Cell extract were made over the course of 2 h to examine Gln3 mobility. Raw data for blots can be found in Supporting information (S1 Raw Images). (B) TUB1-GFP cdc15-2 (YMF3976) cells were grown in YPD and arrested in late anaphase by raising the temperature to 37 °C before the addition of rapamycin to half of the culture. Subsequently, to allow progression through the cell cycle, cells were released in the absence (−) or presence (+) of rapamycin. Samples were taken at the indicated times. Using fluorescence microscopy, the proportion of cells with anaphase spindles in the absence (i) or presence (ii) of rapamycin was investigated. Examples of TUB1-GFP cdc15-2 cells at 120 min after the release at 24 °C are shown in the absence (iii) or presence (iv) of rapamycin. Scale bars indicate 5 μm. (C) cdc15-2 cells (CC2274) were grown in parallel with strains for Fig 2A, but instead or releasing cells at the permissive temperature of 24 °C after the addition of rapamycin like in Fig 2A, cells were maintained at the restrictive temperature of 37 °C in the presence of rapamycin (i). Samples were taken at the indicated times to determine DNA content by flow cytometry analysis (ii) and cell morphology at the end of the experiment (iii). Scale bars indicate 5 μm. (D) cdc14-1 cdc15-2 cells (CC6441) were grown in YPD and arrested in late anaphase by shifting the temperature to 37 °C before the addition of rapamycin to half of the culture (ii). Then, cells were released at 24 °C in the absence (i) or presence (ii) of rapamycin. Samples were taken at the specified times to determine DNA content by FACS analysis. Using light microscopy, we studied cell morphology in the absence (iii) or presence (iv) of rapamycin at the 120 min time point after the release from late anaphase arrest. Scale bars indicate 5 μm. (E) INN1-GFP cdc15-2 (YMF3162) cells were grown as in A. Samples were taken at the indicated times. The proportion of cells with total Inn1-GFP signal in the absence (i) or presence (ii) of rapamycin was determined using fluorescence microscopy. The percentage of cells with either medial rings or spots of Inn1-GFP was calculated too. Examples of cells expressing Inn1-GFP 30 min after the release are shown in the absence (iii) or presence (iv) of rapamycin. Scale bars indicate 5 μm. (F) 3GFP-RAS2 cdc14-1 cells (CC6296) were grown as described in A. Cells are shown at 120 min after the release in the absence (i) or the presence (ii) of rapamycin. Red arrows denote cells where examination of each z-level at the bud neck showed divided cytoplasm and new buds are marked with white asterisks. Scale bars indicate 5 μm. Underlying data for all the graphs can be found in S1 Data file. FACS graphs can be found in the supplementary FACS file (S1 File)., journal.pbio.3002263.s001.pdf, Peer reviewed

Microglial cells are recognized as very dynamic brain cells, screening the environment and sensitive to signals from all other cell types in health and disease. Apolipoprotein D (ApoD), a lipid-binding protein of the Lipocalin family, is required for nervous system optimal function and proper development and maintenance of key neural structures. ApoD has a cell and state-dependent expression in the healthy nervous system, and increases its expression upon aging, damage or neurodegeneration. An extensive overlap exists between processes where ApoD is involved and those where microglia have an active role. However, no study has analyzed the role of ApoD in microglial responses. In this work, we test the hypothesis that ApoD, as an extracellular signal, participates in the intercellular crosstalk sensed by microglia and impacts their responses upon physiological aging or damaging conditions. We find that a significant proportion of ApoD-dependent aging transcriptome are microglia-specific genes, and show that lack of ApoD in vivo dysregulates microglial density in mouse hippocampus in an age-dependent manner. Murine BV2 and primary microglia do not express ApoD, but it can be internalized and targeted to lysosomes, where unlike other cell types it is transiently present. Cytokine secretion profiles and myelin phagocytosis reveal that ApoD has both long-term pre-conditioning effects on microglia as well as acute effects on these microglial immune functions, without significant modification of cell survival. ApoD-triggered cytokine signatures are stimuli (paraquat vs. Aβ oligomers) and sex-dependent. Acute exposure to ApoD induces microglia to switch from their resting state to a secretory and less phagocytic phenotype, while long-term absence of ApoD leads to attenuated cytokine induction and increased myelin uptake, supporting a role for ApoD as priming or immune training factor. This knowledge should help to advance our understanding of the complex responses of microglia during aging and neurodegeneration, where signals received along our lifespan are combined with damage-triggered acute signals, conditioning both beneficial roles and limitations of microglial functions., Peer reviewed

[Introduction]: The Unfolded Protein Response, a mechanism triggered by the cell in response to Endoplasmic reticulum stress, is linked to inflammatory responses. Our aim was to identify novel Unfolded Protein Response-mechanisms that might be involved in triggering or perpetuating the inflammatory response carried out by the Intestinal Epithelial Cells in the context of Inflammatory Bowel Disease., [Methods]: We analyzed the transcriptional profile of human Intestinal Epithelial Cell lines treated with an Endoplasmic Reticulum stress inducer (thapsigargin) and/or proinflammatory stimuli. Several genes were further analyzed in colonic biopsies from Ulcerative Colitis patients and healthy controls. Lastly, we generated Caco-2 cells lacking HMGCS2 by CRISPR Cas-9 and analyzed the functional implications of its absence in Intestinal Epithelial Cells., [Results]: Exposure to a TLR ligand after thapsigargin treatment resulted in a powerful synergistic modulation of gene expression, which led us to identify new genes and pathways that could be involved in inflammatory responses linked to the Unfolded Protein Response. Key differentially expressed genes in the array also exhibited transcriptional alterations in colonic biopsies from active Ulcerative Colitis patients, including NKG2D ligands and the enzyme HMGCS2. Moreover, functional studies showed altered metabolic responses and epithelial barrier integrity in HMGCS2 deficient cell lines., [Conclusion]: We have identified new genes and pathways that are regulated by the Unfolded Protein Response in the context of Inflammatory Bowel Disease including HMGCS2, a gene involved in the metabolism of Short Chain Fatty Acids that may have an important role in intestinal inflammation linked to Endoplasmic Reticulum stress and the resolution of the epithelial damage., Peer reviewed

Strategic Action in Health of the Institute of Health Carlos III Department of Regional Health of the Castilla y Léon Government Consejería de Educación, Junta de Castilla y León, Peer reviewed

Strategic Action in Health of the Institute of Health Carlos III Department of Regional Health of the Castilla y Léon Government Consejería de Educación, Junta de Castilla y León, Peer reviewed