Indirect Immunofluorescence Assay Protocol

Indirect immunofluorescence assays were performed according to previously described protocols (42 (link)). The following primary antibodies were used: mouse anti-HA (Medical & Biological Laboratories Co., Japan), rabbit anti-HSP60, and rabbit anti-TgALD antibodies. Alexa Fluor 488- or Alexa Fluor 594-conjugated secondary antibodies (Life Technologies, USA), Hoechst 33342 (Beyotime, China), and FITC-conjugated Dolichos biflorus agglutinin (DBA-FITC) (Vector Laboratories, USA) were deployed as indicated to visualize the signals. Parasites were imaged with the FV1000 LSCM confocal laser scanning microscope (Olympus, Japan).

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Lyu C., Chen Y., Meng Y., Yang J., Ye S., Niu Z., EI-Debs I., Gupta N, & Shen B. (2023). The Mitochondrial Pyruvate Carrier Coupling Glycolysis and the Tricarboxylic Acid Cycle Is Required for the Asexual Reproduction of Toxoplasma gondii. Microbiology Spectrum, 11(2), e05043-22.

Publication 2023

mouse anti-HA Alexa Fluor 488- or Alexa Fluor 594-conjugated secondary antibodies Hoechst 33342 FITC-conjugated Dolichos biflorus agglutinin (DBA-FITC)

Alexa 594 Alexa fluor 488 Anti antibodies Antibodies Biological Confocal laser scanning microscope Dolichos biflorus agglutinin Fitc Hoechst 33342 Indirect immunofluorescence assays Mouse Parasites Rabbit Vector

Corresponding Organization : Agricultural Genomics Institute at Shenzhen

Other organizations : Humboldt-Universität zu Berlin

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Variable analysis

independent variables

Primary antibodies used: mouse anti-HA, rabbit anti-HSP60, and rabbit anti-Tg ALD antibodies

dependent variables

Visualization of signals using Alexa Fluor 488- or Alexa Fluor 594-conjugated secondary antibodies, Hoechst 33342, and FITC-conjugated Dolichos biflorus agglutinin (DBA-FITC)

control variables

Previously described protocols (42 (link))
Parasites were imaged with the FV1000 LSCM confocal laser scanning microscope (Olympus, Japan)

controls

No positive or negative controls were explicitly mentioned in the provided information.

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