Project description:We performed MNase-ChIP seq with deep digestion to identify the MuvB associated nucleosomes in WT and LIN37 -/- HCT116 cells in G1 arrested and cycling cells. Strep-LIN9 as well as a mutant of the protein (Strep-4xLIN9: R174A/R175A/F180A/F181A) which is unable to associate with LIN37 or RbAP48 were transiently transfected in WT and LIN37 -/- HCT116 cells. At 24 hours after transfection, cells were treated with Nutlin-3a to induce G1 arrest; cells were harvested 48 hours following treatment. For several samples, Nutlin-3a treatment was omitted to maintain cycling conditions. Cells were then cross-linked with formaldehyde, lysed and treated with micrococcal nuclease. Chromatin fragments were precipitated with Streptactin-XT magnetic beads after which DNA was purified and libraries were prepared and sequenced using NovaSeq 6000 platform in 150bp paired-end mode. We find that LIN9 precipitates to +1 nucleosomes of target genes in arrested HCT116 and LIN37 -/- cells. The precipitation of this nucleosome is diminished in cycling cells as well as in arrested cells transfected with the Strep-4xLIN9 mutant.
Project description:Phosphoproteome of RA264.7 macrophages 30m post infection with quorum sensing ON or OFF or both Streptococcus pyogenes (Group A Strep), or uninfected. Available are peak lists, .raw files, and processed and quantified results analyzed via Comet.
2025-01-22 | MSV000096914 | MassIVE
Project description:Strep. gordonii and Oral Biofilm Formation
| PRJNA882118 | ENA
Project description:Strep. gordonii and Oral Biofilm Formation
Project description:Corynebacterium glutamicum GlxR is a homolog of the cAMP receptor protein. Although over 200 GlxR binding sites in the C. glutamicum genome are predicted in silico, studies on the GlxR physiological function have been hindered by the severe growth defects of a glxR mutant. This study comprehensively identified the GlxR regulon by chromatin immunoprecipitation in conjunction with microarray (ChIP-chip) analyses. In total, 209 regions were detected as in vivo GlxR binding sites. Moreover, ChIP-chip analyses showed that GlxR was still able to interact with its target sites in a deletion mutant of cyaB, the sole adenylate cyclase gene in the genome, even though binding affinity was markedly decreased. To identify the direct GlxR targets, we immunoprecipitated DNA from a strain expressing a Strep-tag II-tagged GlxR-protein using an anti-Strep-tag II antibody. To investigate effect of depletion of cAMP by deletion of the cyaB gene, which encodes the sole adenylate cyclase in C. glutamicum, on GlxR binding in vivo, we immunoprecipitated DNA from a cyaB deletion strain expressing a Strep-tag II-tagged GlxR-protein using an anti-Strep-tag II antibody. Three or more independent biological replicates were generated in both cases.
Project description:The aim of this study was to uncover cell cycle dependent chromatin binding of H2A.Z and its histone chaperones. U2OS cells were stably transfected with Twin-Strep-tagged H2A.Z, ANP32e, or YL1 constructs, and WT cells were used as a negative/background control. U2OS cells were synchronised in G1 or G2-M phase using hydroxyurea or nocodazole, respectively, and protein-DNA complexes were isolated by affinity purification.
Project description:Gene expression in the obligatory aerobic acetic acid bacterium Gluconobacter oxydans was shown to respond to oxygen limitation, but the regulators involved are unknown. In this study, we analyzed the function of a transcriptional regulator named GoxR, which belongs to the FNR family. Here, we applied ChAP-seq analysis with a strep-tagged GoxR version to identify binding sites of this regulator in the genome of G. oxydans.
Project description:In this dataset, we included sequencing data of total and ribosome protected fragments obtained from PATU-8902 cell lines grown in DMEM (2.78mM glucose, 4mM glutamine, 1mM pyruvate) + 10%dialyzed FBS + 1% Pen/Strep with or without 400uM Ser and 400uM Gly for 24 hours.
Project description:Comparison of gene expression profiles of the GL261 cell line (a murine glioma model) grown in duplicate in two different types of media. AC samples where grown in DMEM supplemented by 20% FBS, 5 U/ml pen/strep and 4 mM L-glutamine. NS samples were grown in DMEM/F12 (50/50) supplemented with 2 U/ml pen/strep, 1 ug/ml fungizone, 1x B27, 20 ng/ml bFGF, 20 ng/ml EGF, 20 ng/ml LIF and 5 ug/ml heparin. We have reason to believe the NS media enhances cell de-differentiation.