Human cerebral microvascular endothelial cells (HCMECs) were infected with extraintestinal pathogenic Escherichia coli (ExPEC) strain RS218 at a multiplicity of infection (MOI) of 1 for 3 hours. Proteomic analysis was performed to evaluate host cellular responses. Infected samples were compared to uninfected controls (MOCK) to identify differentially expressed proteins associated with ExPEC infection.