Author: He, Cheng; Kong, Ling; Zhou, Lijuan; Xia, Jing; Wei, Haixia; Liu, Min; Peng, Hongjuan
Title: Host Cell Vimentin Restrains Toxoplasma gondii Invasion and Phosphorylation of Vimentin is Partially Regulated by Interaction with TgROP18 Document date: 2017_9_5
ID: z9mg639h_39
Snippet: Phos-tag acrylamide (Wako Chemicals) based gel analysis was carried out to detect phosphorylated vimentin. Cos7 cells were infected with T. gondii RH or RH Δrop18 strains at a MOI of 3 for different periods of time (0, 1, 2, 6, 12, 24, 36, and 48h). Cells were then lysed with cell lysis buffer (TransGen Biotech, DE101) and the extracts were loaded on to 10% polyacrylamide gel (separating gel) containing 50μM Phos-tag acrylamide (Wako Chemicals,.....
Document: Phos-tag acrylamide (Wako Chemicals) based gel analysis was carried out to detect phosphorylated vimentin. Cos7 cells were infected with T. gondii RH or RH Δrop18 strains at a MOI of 3 for different periods of time (0, 1, 2, 6, 12, 24, 36, and 48h). Cells were then lysed with cell lysis buffer (TransGen Biotech, DE101) and the extracts were loaded on to 10% polyacrylamide gel (separating gel) containing 50μM Phos-tag acrylamide (Wako Chemicals, AAL-107). As vimentin expression levels changed with the time post T. gondii infection, the amount of cell lysate protein loaded for SDS-PAGE was adjusted to ensure a consistent level of total vimentin protein.
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