Quantitative Proteomics Using RIME and Dimethyl-Labelling
RIME and stable isotope dimethyl-labelling were performed, as previously described17 (link),60 (link),61 (link). Briefly, cells were labelled with the medium and light isotope reagent. Labelled samples were pooled at an approximate 1:1 ratio, dried down and fractionated using 12 cm IPG strip pH 3-1062 (link). RIME and dimethyl-label fractions were desalted (SUM SS18V, The Nest Group Inc) and run through LC-MS/MS using LTQ Velos Orbitrap MS. Raw data for RIME and dimethyl-labelling were analysed using MaxQuant 1.5.1.062 (link),63 (link). Statistical analysis was performed using Perseus software (version 1.6.1.3)64 (link). Raw data have been deposited to the ProteomeXchange Consortium via PRIDE partner repository with the dataset identifier PXD004648 (MCF-7) and PXD004807 (SUM44).
Montaudon E., Nikitorowicz-Buniak J., Sourd L., Morisset L., El Botty R., Huguet L., Dahmani A., Painsec P., Nemati F., Vacher S., Chemlali W., Masliah-Planchon J., Château-Joubert S., Rega C., Leal M.F., Simigdala N., Pancholi S., Ribas R., Nicolas A., Meseure D., Vincent-Salomon A., Reyes C., Rapinat A., Gentien D., Larcher T., Bohec M., Baulande S., Bernard V., Decaudin D., Coussy F., Le Romancer M., Dutertre G., Tariq Z., Cottu P., Driouch K., Bièche I., Martin L.A, & Marangoni E. (2020). PLK1 inhibition exhibits strong anti-tumoral activity in CCND1-driven breast cancer metastases with acquired palbociclib resistance. Nature Communications, 11, 4053.
Other organizations :
Université Paris Sciences et Lettres, Institut Curie, Institute of Cancer Research, École Nationale Vétérinaire d'Alfort, Oniris, Centre de Recherche en Cancérologie de Lyon
RIME and stable isotope dimethyl-labelling were performed
dependent variables
Labelled samples were pooled at an approximate 1:1 ratio
Fractionated using 12 cm IPG strip pH 3-10
RIME and dimethyl-label fractions were desalted and run through LC-MS/MS using LTQ Velos Orbitrap MS
Raw data for RIME and dimethyl-labelling were analysed using MaxQuant 1.5.1.0
Statistical analysis was performed using Perseus software (version 1.6.1.3)
control variables
Cells were labelled with the medium and light isotope reagent
positive controls
None specified
negative controls
None specified
Annotations
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