Host Cell Vimentin Restrains Toxoplasma gondii Invasion and Phosphorylation of Vimentin is Partially Regulated by Interaction with TgROP18

He, Cheng; Kong, Ling; Zhou, Lijuan; Xia, Jing; Wei, Haixia; Liu, Min; Peng, Hongjuan

Resulting parasite-infected monolayers were fixed with 4% formaldehyde for 15 min, permeabilized with 0.1% Triton X-100 for 20 min at room temperature, and blocked in 10% FBS for 15 min [55] . Samples were incubated with primary antibodies overnight at 4°C, and then washed in PBS with gentle shaking at RT three times for 5min, and incubated with secondary antibodies (goat anti-mouse IgG or goat anti-rabbit IgG) conjugated with FITC, Texas Red ® , or TRITC, diluted 1:200. Following staining, slides were again washed with PBS (as described above), and then rinsed with ddH 2 O. Air-dried slides were mounted with DAPI Fluoromount ® (Southern Biotech). Samples were visualized and images generated using an Olympus FLUOVIEW FV1000 confocal laser scanning microscope or by fluorescence microscopy.