Project description:Here, we used APEX2 proximity labelling technique coupled with mass spectrometry to define Cveolin-1 interactome at steady state and after hypo-osmotic shock. We created a stable cell line expressing Caveolin1-APEX2-EGFP in human retinal pigmented epithelial cells (RPE-1). Hypo osmotic shock was performed accordingly to Sinha et al, 2011. Briefly, cells were incubated in 30mOsm medium for 5 minutes (Hypo condition). To analyse the ability to recover, after the osmotic shock cells were allowed to recover in isotonic medium for 30 minutes (REC condition). Cells grown in iso-tonic medium were used as normal non treated condition (NT). RPE-1 cells expressing APEX2-EGFP in the cytoplasm (NES-APEX2-EGFP) was used as control to exclude highly abundant cytosolic proteins.
Project description:To better understand how yeast adapt and respond to sequential stressors, an industrial yeast strain, URM 6670 (also known as BT0510), which is highly flocculent, tolerant to ethanol, osmotic and heat shock stresses, was subjected to three different treatments: 1. osmotic stress followed by ethanol stress, 2. oxidative stress followed by ethanol stress, 3. glucose withdrawal followed by ethanol stress. Samples were collected before the first stress (control), after the first stress and after the second stress (ethanol). RNA was extracted and analyzed by RNAseq.
Project description:S. cerevisae cells were exposed to different series of mild stresses. Stress type include heat shock, oxidative and osmotic stresses. Microarrays were used to follow the genome-wide transcriptional response to the stresses and to identify genes that can underlie the cross protection phenotype between heat shock and oxidative stress. Experiment Overall Design: Cell sample at different time points after stress application were used for RNA extraction and hybridization on Affymetrix microarrays.
Project description:Yeast cells can be affected during their growth to several stress conditions. One of the most known and characterised is the osmotic stress and most of the studies about osmotic sterss response in yeast have been focused on salt or sorbitol stress. However, during yeast growth in industrially relevant processes (for instance throughout alcoholic fermentation on the must to produce alcoholic beverages) the osmotic stress is mainly due to the high sugar(in particular glucose) concentration (200-250 g/L).
