MassIVE MSV000090607

Partial Public PXD037794

Proteome of senescent human lung fibroblasts and Panc-1 cells

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Description

Proteomic analysis of proliferating and senescent normal healthy lung fibroblasts (NHLFs) and Panc-1 cells. Senescence was induced using 5 uM Palbociclib for 6 days. Cells were lysed in SDS lysis buffer (2% SDS in 100 mM TEAB), boiled (15 min, 95 °C, 500 rpm) and sonicated in a sonication bath (10 min). DNA was sheared with Benzonase® Nuclease (Merck Millipore) and 2 mM MgCl2 at 37 °C (30 min, 500 rpm). Reversibly oxidized cysteines were reduced with 10 mM TCEP followed by alkylation with 20 mM IAA (each step 30 min, 22 °C, 600 rpm, in the dark). Proteins were acetone precipitated and the protein pellet was resuspended in 100 µL 100 mM TEAB. Proteins were digested overnight with LysC/trypsin (1:50 protease:protein ratio, 37 °C, 600 rpm) Peptides were desalted by solid-phase extraction using Oasis HLB cartridges (10 mg; Waters), resuspended in 3% ACN/0.1% TFA and analysed on an Orbitrap Eclipse Tribrid mass spectrometer in combination with the FAIMS Pro Interface (Thermo Scientific). [doi:10.25345/C5KH0F391] [dataset license: CC0 1.0 Universal (CC0 1.0)]

Keywords: senescence, NHLF, PANC-1, fibroblasts, lung, palbociclib

Contact

Principal Investigators:
(in alphabetical order) 		Karim C. El Kasmi, Boehringer Ingelheim Pharma GmbH & Co. KG, Biberach an der Riss, Germany
Submitting User: egriesser

Publications

Schloesser D, Lindenthal L, Sauer J, Chung KJ, Chavakis T, Griesser E, Baskaran P, Maier-Habelsberger U, Fundel-Clemens K, Schlotthauer I, Watson CK, Swee LK, Igney F, Park JE, Huber-Lang MS, Thomas MJ, El Kasmi KC, Murray PJ.
Senescent cells suppress macrophage-mediated corpse removal via upregulation of the CD47-QPCT/L axis.
J Cell Biol. Epub 2022 Dec 2.

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