Journal: Frontiers in Veterinary Science

Article Title: AMPK and CAMKK activation participate in early events of Toxoplasma gondii -triggered NET formation in bovine polymorphonuclear neutrophils

doi: 10.3389/fvets.2025.1557509

Figure Lengend Snippet: Toxoplasma gondii -induced DNA release in bovine PMN is dependent on ERK and SOCE signalling. Bovine PMN ( n = 3) were pre-treated for 30 min with UO126 (50 μM), LY294002 (1 μM) or 2-APB (50 μM) before the addition of T. gondii (1:4 PMN: tachyzoites ratio). After 4 h of co-incubation, the Picogreen-derived fluorescence, corresponding to extracellular DNA amount, was determined in a plate reader. DNAse I (90 U) was added after the 4 h of incubation in the corresponding experiments to confirm the DNA-nature of the emitted fluorescence. Bars represent the mean ± SD. p -values were calculated by applying an ANOVA test followed by a Dunnet multiple comparison test.

Article Snippet: DNA was quantified via PicoGreen-based dsDNA quantitation reagent (5 μM, Invitrogen)-derived fluorescence intensities being assessed at an excitation wavelength of 500 nm and emission wavelength 525 nm using an automated plate monochrome reader (Varioskan Flash; Thermo Scientific) after 4 h of co-culture.

Techniques: Incubation, Derivative Assay, Fluorescence, Comparison