NIH 3T3 AHA p-c, SILAC p-c, inhibitor, and enrichment control experiments

ABSTRACT: We developed a pulse-chase method in where fully SILAC (heavy, medium-heavy and Light) labelled cells were pulsed with Azidohomoalanine (AHA) and then chased for different length of times. These experiments were performed with 0, 1, 2, 4, 8, 16 and 13 h of chase. Data from 3 biological replicates are provided. Each replicate contains data from 3 experiments (0, 1, 2 and 0, 4, 8 and 0, 16, 32 hours chase) in where the 0 h time point is always heavy labelled followed by the medium and light labelled time points. In addition, AHA p-c experiments were performed using only 0, 4 and 8 h chase times in combination with different inhibitor combinations (MG132, Actinomycin D and Wortmanninin + Bafilomycin A1) or control (DMSO). Also, a control enrichment data set was created. Here heavy SILAC labelled cells were pulsed with AHA before being mixed with light cells that were not pulsed with AHA. This was followed by the click reaction and enrichment of AHA containing proteins. Label-swap experiments were also performed. Finally, a SILAC p-c data set is provided. Here light cells were pulsed with heavy (Arg10, Lys8) amino acids and then split in two. One half of the cells was chased in medium (Arg6, Lys4) amino acids and the other part was directly frozen. Label-swap experiments and experiments using different pulse and chase times were also performed. All provided datasets are from mouse fibroblast cells (NIH 3T3).

INSTRUMENT(S): Q Exactive

ORGANISM(S): Mus Musculus (mouse)

TISSUE(S): Fibroblast

SUBMITTER: Erik McShane  

LAB HEAD: Matthias Selbach

PROVIDER: PXD004929 | Pride | 2016-10-06

REPOSITORIES: pride