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To determine the secretion of CPs, 1 × 106 trophozoites were suspended in 500 µl TYI_secretion medium. After 0 min, 1 h, 2 h, and 3 h of incubation, the amoebae were sedimented and the supernatant was removed to measure CP activity. To determine the total CP activity, soluble extracts of 1 × 106 trophozoites/500 µl were generated. Secretion was outlined as the percentage of total activity. Each time point was measured at least two times in duplicate.
