Project description:To determine the interactome of RNA Polymerase II (RNAPII), HA-tagged RPB3 was stably expressed in U2OS cells. The cells were harvested followed by the native stepwise isolation of chromatin-associated RNAPII complexes. HA-RPB3 was immunoprecipitated and interacting proteins analyzed by quantitative mass spectrometry. U2OS cells expressing no HA-tagged protein were used for comparison.
Project description:The purpose of this experiment was to identify the genes bound by Ndt80 in P. pastoris when constitutively expressed. Ndt80 was tagged with c-myc, the native Ndt80 promoter was replaced with the E. gossypii Tef1 promoter and the protein was immunoprecipitated with a c-myc antibody. Cells were grown unti log-phase in YPD. Each experiment was repeated twice and sequenced on an Illumina HiSeq 2500.
Project description:The purpose of this experiment was to identify the genes bound by Ndt80 in S. cerevisiae during meiosis and sporulation. Ndt80 was tagged with c-myc and the protein was immunoprecipitated with a c-myc antibody. Cells were grown in liquid YPA (2% Peptone, 2% Potassium Acetate, 1% Yeast Extract) at room temperature for 22 hours. Each experiment was repeated twice and sequenced on an Illumina HiSeq 4000.
Project description:The purpose of this experiment was to identify the genes bound by the two Ndt80 paralogs in S. stipitis when constitutively expressed. Both paralogs were tagged with c-myc, the native Ndt80 promoter was replaced with the E. gossypii Tef1 promoter and the protein was immunoprecipitated with a c-myc antibody. Cells were grown unti log-phase in YPD. Each experiment was repeated twice and sequenced on an Illumina HiSeq 2500.
Project description:The purpose of this experiment was to identify the genes bound by Ndt80 in S. cerevisiae when constitutively expressed, for the native Ndt80 as well as a heterologous Ndt80 from Pichia pastoris. For the ChIPs on the native Ndt80s, Ndt80 was tagged with c-myc; for the heterologous ChIP P. pastoris Ndt80 was integrated into the S. cerevisiae genome and tagged with c-myc. In both cases, the native Ndt80 promoter was also replaced with an A. gossypii Gal1 promoter and the protein was immunoprecipitated with a c-myc antibody. Cells were grown overnight in SRaffinose until log-phase, then in SRaffinose + 1.7% Galactose for 5 hours. Each experiment was repeated twice and sequenced on an Illumina HiSeq 4000.
Project description:The purpose of this experiment was to identify the genes bound by Ndt80 in K. lactis when constitutively expressed. Ndt80 was tagged with c-myc, the native Ndt80 promoter was replaced with the K. lactis Gal1 promoter and the protein was immunoprecipitated with a c-myc antibody. Cells were grown overnight in SRaffinose until log-phase, then in SRaffinose + 1.7% Galactose for 5 hours. Each experiment was repeated twice and sequenced on an Illumina HiSeq 4000.
Project description:Myc-tagged FBP1-WT, Myc-tagged FBP1-G164D, Myc-tagged FBP1-F194S or empty vector were transfected into FBP1-KO HepG2 cells. FBP1 complexes were immunoprecipitated with anti-Myc beads and then separated by SDS‒PAGE. Bands were removed from the gel, and trypsin-digested samples were subjected to liquid chromatography-tandem mass spectrometry (LC-MS/MS) using an Orbitrap Velos Pro (Thermo Scientific). MS/MS dataset analysis was performed using the Mascot software program (Matrix Science).
Project description:The purpose of this experiment was to identify the genes bound by the two Ndt80 paralogs in C. albicans when constitutively expressed, and for Ndt80B when endogenously expressed. Both paralogs were tagged with c-myc, and the protein was immunoprecipitated with a c-myc antibody. For constitutive expression, each native Ndt80 promoter was replaced by the C. albicans Tdh3 promoter. Cells were grown unti log-phase in YPD. Each experiment was repeated twice and sequenced on an Illumina HiSeq 2500.
Project description:The purpose of this experiment was to determine the genes directly regulated by Ste12 in K. lactis. The experiment was performed in a cells to determine if the a-specific genes were bound by Ste12. Ste12 was tagged with c-myc and was immunoprecipitated with a c-myc antibody. Cells were starved in SD media lacking phosphate for 2 hours, then treated with 10µg/mL K. lactis alpha factor for 2 hours before harvesting. A control untagged strain was processed in parallel. The experiment was repeated 3 times, and sequenced on an Illumina HiSeq 2500.
Project description:Myc-tagged N-terminal portion of TOC1 or TOC1 mutant (S194A, S201A, T204A) were expressed in tobacco leaves and then immunoprecipitated. The immunoprecipitated samples were separated on SDS-PAGE gels, and then phosphorylation sites of TOC1 were identified from gel bands.