p-IκBα localizes to the parasitophorous vacuolar membrane of RH-ERP, but not RH-JSR or GT1, and is partially dependent on GRA2. (A) Human foreskin fibroblasts were infected with type I strains for 24 hours, fixed with formaldehyde and stained with p-IκBα (red), GRA7 (green) and Hoechst (blue). Pictures are representative of at least three experiments. (B) Quantification of p-IκBα recruited to the PVM of RH-ERP, RH∆gra2, RH-JSR and GT1. The intensity of p-IκBα recruited to the PVM was quantified in at least 5 cells per condition. The graph shows the average from three independent experiments, with levels showing average p-IκBα recruitment quantification, and the error bars represent standard error. * p-value < 0.01, Student’s t test. (C) HFFs were infected with RH-GFP and GT1 for 24 hours, fixed with formaldehyde and stained with p-IκBα (red), GRA7 (green) and Hoechst (blue). Arrows point to GT1 vacuoles. (D) Mouse embryonic fibroblasts were infected with RH-ERP for 30 hours, fixed with formaldehyde and stained with p-IκBα (red), GRA7 (green) and Hoechst (blue). Arrows point to the intervacuolar PVM extension.