Multilayered-proteomics analysis of insulin signaling in insulin sensitive and resistant HepG2 IGF1R KO cell line

ABSTRACT: This is a mass spectrometry (MS)-based proteomics dataset, generated to study insulin signaling in a hepatocellular cell model (HepG2 insulin-like growth factor knock-down), at the IR interactome, phos-phoproteome, and proteome level. To induce insulin sensitivity and resistance in HepG2 IGF1R KO cells, the following protocol, established based on a previously published study by Dall’Agnese et al. (https://doi.org/10.1038/s41467-022-35176-7), was used. The day after cell seeding, culture medium was changed to serum-free low glucose DMEM for 48 hours. Following the serum washout, cells were treated for 48 hours in low glucose DMEM with 1.25 % HSA (human serum albumin) and either a phys-iologic level of 0.1 nM insulin and a pathologic level of 3 nM insulin, to make the cells either sensitive or resistant to insulin. Insulin sensitive and resistant HepG2 IGF1R KO cells, were subjected to an insulin wash-out over 35 minutes, with 7 media exchanges. Afterwards, the cells were stimulated for 5 minutes with varying insulin concentrations (0, 0.1, 3, or 100 nM) in low glucose DMEM with 1.25% HSA. Note that 0.1 nM is named 100 pM in the files. We performed 5 biological independent experiments, which were used to study the IR interactome, phosphoproteome, and a reference single-shot proteome. These cells were lysed in a co-immunoprecipitation (IP)-lysis buffer, preserving protein-interactions. Additional-ly, we generated SDS-based label-free DIA-MS single-shot proteome dataset, from four independent experiments of the insulin sensitive and resistant HepG2 IGF1R KO cells, directly after the insulin-resistance inducing procedure.

INSTRUMENT(S):

ORGANISM(S): Homo Sapiens (human)

TISSUE(S): Hepatocyte

SUBMITTER: Sarah Jørgensen  

LAB HEAD: Jesper Velgaard Olsen

PROVIDER: PXD047011 | Pride | 2025-05-06

REPOSITORIES: Pride