Journal: bioRxiv

Article Title: An ambiguous N-terminus drives the dual targeting of an antioxidant protein Thioredoxin peroxidase (TgTPx1/2) to endosymbiotic organelles in Toxoplasma gondii

doi: 10.1101/562587

Figure Lengend Snippet: A, B) Microscopic images of T. gondii parasites showing localization of an HA tagged TgTPx1/2 (red) with a mitochondrial marker SPTP-SOD2-GFP (green) and an apicoplast marker ACP (green). C) A Western blot analysis for total cell lysate of parasites stably expressing TgTPx1/2-HA using anti-HA antibodies. Full-length Western blot is represented in Supplementary Fig. S3. D, E) Fluorescence images of parasites stably expressing TgTPx1/2N 1-50 -EGFP (green) with a mitochondrial marker SPTP-SOD2-DsRed (red) and apicoplast maker, ACP (red). Scale bar, 2µm. Numbers (in black) at the top right corner of the DIC+Merge panel indicates the number of parasites inside the vacuole.

Article Snippet: The proteins were transferred to PVDF membrane, blocked for an hour with 5% BSA-Tris Buffered Saline (TBS) and probed with primary antibodies anti-HA (C29F4) Rabbit monoclonal antibody (mAb) (Cell Signaling Technology®) or mouse raised anti-GFP antibodies (Roche) for 2 hours.

Techniques: Marker, Western Blot, Stable Transfection, Expressing, Fluorescence

Journal: bioRxiv

Article Title: An ambiguous N-terminus drives the dual targeting of an antioxidant protein Thioredoxin peroxidase (TgTPx1/2) to endosymbiotic organelles in Toxoplasma gondii

doi: 10.1101/562587

Figure Lengend Snippet: A) SignalP 3.0-HMM and MitoProt scores for the wildtype TgTPx1/2 and the mutants TgTPx1/2(R24A) and TgTPx1/2(L17A,L27A). Immunofluorescence images of the parasites expressing B, C) TgTPx1/2(R24A) (red/green) and D, E) TgTPx1/2(L17A,L27A) (green/red). Here ACP is used to label the apicoplast (green) while Mitotracker Red (red) is used for labelling the mitochondrion of T. gondii . Scale bar, 2µm. Numbers (in black) at the top left corner of the DIC panel indicates the number of parasites inside the vacuole. F) A Western blot analysis for the whole parasite lysates of wildtype TgTPx1/2, TgTPx1/2(R24A) and TgTPx1/2(L17A,L27A) using anti-HA antibodies. Full-length Western blot is represented in Supplementary Fig. S3.

Article Snippet: The proteins were transferred to PVDF membrane, blocked for an hour with 5% BSA-Tris Buffered Saline (TBS) and probed with primary antibodies anti-HA (C29F4) Rabbit monoclonal antibody (mAb) (Cell Signaling Technology®) or mouse raised anti-GFP antibodies (Roche) for 2 hours.

Techniques: Immunofluorescence, Expressing, Western Blot