Project description:In order to identify the effects of starvation on the MEFs wt trascriptome, we performed Affymetrix Gene-Chip hybridization experiments for the starved cells Transcriptome analysis of the starved MEFs wt cells For the analysis on the starved MEFs wt cells, total RNA was extracted; RNA extracted from MEFs wt cells grown in Normal Medium was used as control
Project description:In order to identify the effects of starvation on the MEFs wt trascriptome, we performed Affymetrix Gene-Chip hybridization experiments for the starved cells Transcriptome analysis of the starved MEFs wt cells
Project description:Mouse embryonic fibroblasts can be used to condition basis human ES cell medium to allow feeder-free growth. To investigate the impact of factors released by the MEFs on gene expression in hES cells, global gene expression was analysed in cells grown in MEF-conditioned medium as compared to cells grown in a chemically defined one. Keywords: Media comparison
Project description:(1)In vivo SILAC analyses of mouse embryonic fibroblasts (MEFs, triple labeling) were performed comparing phosphorylation events. ULK1 double knock out MEFs (VC) were compared to double knock out MEFs expressing human ULK1 (RE) in fed (DMEM) and starvation (HBSS) conditions. (2) In vivo SILAC analyses of A549 cells (triple labeling) were performed comparing phosphorylation events (195 raw files labeled ""A549""). A549 cells in fed conditions (DMEM) were compared to starved cells (HBSS) and cells treated with rapamycin (Rapa).
Project description:Hat1 WT and Hat1 KO MEFs were grown on both galactose and glucose. Abundance of acetylated peptides was compared between the samples.
Project description:Understanding the regulation of lysosomal proteolytic/hidrolytic capacity has been recently advanced in a huge manner by the discovery of the lysosome-mTOR-TFEB pathway connecting nutrient sensing, autophagy and de novo lysosome generation. But, how lysosomes can adjust their proteolytic/hidrolytic capacity to meet varying conditions (more or less substrate) under normal fed conditions is not known. We have used AEP as a proxy to understand how lysosomes can compensate the lack of a key lysosomal protease to meet the requirements under physiological, fed conditions. To do this, we did compare cytoplasmic, membrane and nuclear fractions of WT and AEP MEFs that were expanded in SILAC media. AEP knock-out mice show pale kidney, with abnormal proliferation of the epithelial proximal tubular cells. In an attempt to understand the role of AEP in the kidney, we did compare kidney obtained from WT and AEP mice using kidneys from WT mice that were fed with SILAC food. Also, in an attempt to reproduce the changes observed in the absence of AEP in kidney and MEFs, we treated WT MEFs grown in SILAC media with MVO26630, specific inhibitor of AEP, for 12 and 48 hours.
Project description:v-ATPase Vo and V1 domains were isolated via Co-IP from MEFs expressing HA-Atp6V1B2 and FLAG-Atp6VoaA3 using HA- and FLAG-tag antibodies. Treatment groups were full medium (control), amino acid starvation (60 min) and restimulation (30 min). Cells were grown in SILAC medium for 3 passages.
Project description:In order to identify the effects of starvation on the PPP3R1 cell line trascriptome, we performed Affymetrix Gene-Chip hybridization experiments for the starved cells Transcriptome analysis of the starved PPP3R1 cell line For the analysis on the starved PPP3R1 cell line, total RNA was extracted; RNA extracted from PPP3R1 cell line grown in Normal Medium was used as control
Project description:Primary lung fibroblasts were grown to confluence in DMEM 10% FBS, serum starved in DMEM 0.5% FBS for 18 hours. Cells were then treated in the presence or absence of 100 nM ET-1 in the same medium for an additional 4 hours. Keywords: repeat sample
