Fig. 3
From: Neuroglobin regulates autophagy through mTORC1/RAPTOR/ULK-1 pathway in human neuroblastoma cells
Analysis of the NGB interactome. (A) Protein complexes from NGB-FLAG (n = 3) and GFP-FLAG (n = 3) SH-SY5Y cell samples were enriched by co-IP and the specific elution of FLAG-tagged complexes by competition with FLAG peptide was assessed by SDS-PAGE. IP = immunoprecipitation, WCE = whole cell extracts. (B) Volcano plot shows the results of proteomic analysis of NGB-FLAG and GFP-FLAG protein complexes after identification and statistical selection of putative NGB interactors (blue circles). (C) Putative NGB interactors were used to construct a full PPI network with (D) its corresponding functional enrichment. (E) Venn analysis was performed to analyze the overlap between full NGB’s interactome and other published NGB’s interactomes data. (F) High-confidence NGB interactors were selected after CRAPome analysis and used to build a refined network with (G) its corresponding functional enrichment. (H) The same list of high-confidence NGB interactors was used to build a refined STRING network. (I) PPI prediction analysis was performed to calculate the value of predictive interaction between NGB and RPTOR. The structures of the two potential interacting proteins were reported as non-interacting chains, with a likelihood ratio (LR)= -2.455.
