Fig. 2. ERO1 loss impairs the secretion of HIF-1-dependent and angiogenesis-relevant targets.

A Distribution of total intracellular peptides of WT and ERO1 KO MDAMB231^m under normoxic and hypoxic conditions, using nLC-MS/MS proteomic analyses. The redox state of cysteines is reported as free thiol if the cysteines are alkylated by N-ethylmaleimide (NEM) and present in a reduced form; or as disulfide-bond if the cysteines are alkylated by carbamidomethylation (IAA) and are present in an oxidized form. B Distribution of intracellular proteins containing differently IAA/NEM-labeled peptides in WT and ERO1 KO MDAMB231^m. C Distribution of secreted proteins (secretome by nLC-MS/MS proteomic analysis) containing differently IAA/NEM-labeled peptides in WT and ERO1 KO MDAMB231^m. Proteins containing the differently IAA-labeled peptides and identified in WT are shown as the gene name. D Table reporting the fold change of HIF-1 target secreted proteins as gene name (MetaCore analytical suite version 19.4) in WT and ERO1 KO MDAMB231^m in hypoxic conditions by secretome proteomics (nLC-MS/MS proteomic analyses) (N = 3). “Only WT” refers to protein identified only in the WT group. * highlighted secreted proteins whose fold change was significantly different between ERO1 KO vs. WT cells (Wilcoxon Mann–Whitney test, p < 0.05, JMP pro13). E Bar graphs presenting levels of angiogenic cytokines detected by a human angiogenic array in the secretome of WT and ERO1 KO MDAMB231^m undergoing hypoxia for 48 h (N = 3). The reference spot was set to 1. The cytokines with a value of WT above 0,1 are shown.