Project description:Transcriptomics of NIH-3T3 cells treated with typical or atypical RARa modulators to evaluate genetic changes induced by chemical modification of RARa signalling
Project description:NIH/3T3 cells were cultured at least 2 weeks in light medium: DMEM for SILAC (Thermo scientific) supplemented with PS, dialyzed FBS (Thermo scientific), (light) L-Lysine-2HCl (0.666 mM), (light) L-Arginine-HCl (0.399 mM)44, and L-Proline (200 mg/L). Before switching from light to heavy medium, cells were treated with 0.05 µg/mL CHX or equivalent dilution of DMSO in 6-well plates. At timepoint 0 h, we replaced light medium with pre-warmed heavy medium: DMEM for SILAC (Thermo scientific) supplemented with PS, dialyzed FBS (Thermo scientific), (heavy) 13C6 15N2 L-Lysine-2HCl (0.666 mM), (heavy) 13C6 15N4 L-Arginine-HCl (0.399 mM), and L-Proline (200 mg/L) for NIH/3T3.
Project description:Co-IP/MS of Gli3 from NIH/3T3 cells treated with the smoothened agonist SAG fractionated into nuclear and cytoplasmic fractions. 10091, 10092, 10093, 10095, 10096 are nuclear fraction samples, 10094, 10097 are cytoplasmic fraction samples.
Project description:In this study, we compared the transcriptional profile of NIH-3T3 cells that are either wild-type (WT) or Nrf1 knockout (Nrf1-KO). We treated these cells with carfilzomib for 6 hours or 24 hours and performed RNA-sequencing with these samples.
Project description:Analysis of Immediate Early Response 2 (Ier2)-inducible NIH 3T3 cells after Ier2 induction with RheoSwitch ligand RSL-1. Results provide insight into the function of Ier2 in NIH 3T3 mouse embryonal fibroblasts. Immediate early genes, including Ier2, are rapidly induced in quiescent cells by proliferation and migration-inducing stimuli. Microarray gene expression profiling was performed to identify differentially expressed genes following overexpression of Ier2 in NIH 3T3-Ier2 inducible cells after 24 hour induction of Ier2.
Project description:Expression data from NIH-3T3 cells treated with mock, 100 U/ml IFN alpha or 100 U/ml gamma for 1 or 3h on nt-RNA labeled for 30-60 min at different times of interferon treatment; Differential gene expression caused by IFN alpha or gamma was analyzed in newly transcribed RNA (nt-RNA) of NIH-3T3 cells treated for 1 or 3 h. RNA was labeled for 30 to 60 min and separated from total cellular RNA (tc-RNA) following Trizol RNA preparation and thiol-specific biotinylation; We used microarrays to analyze the effects of IFNalpha and gamma treatment in newly transcribed RNA (nt-RNA) Experiment Overall Design: NIH-3T3 cells (5th to 15th passage after thawing) were split 24 h before start of the experiment. When starting the experiment 80% confluency was reached. The experiment was started by applying fresh, prewarmed, CO2-equilibrated medium containing either mock, 100U/ml IFN alpha or 100 U/ml IFN gamma. Labeling was either started simultaneously, 30 or 150 minutes later.