Fig. 5

Transcriptional changes in AcSir2 during encystation and the effects of AcSir2 overexpression on the encystation of A. castellanii. a Changes in the expression of AcSir2 during the encystation of A. castellanii. Trophozoites and encysting cells 24, 48 and 72 h after the induction of encystation were examined in terms of AcSir2 transcriptional changes using qRT-PCR. Acanthamoeba actin was used as an internal control. b, c Effects of AcSir2 overexpression on the encystation of A. castellanii. Vector control (left) and AcSir2-overexpressing trophozoites (right) were transferred into encystation medium, incubated for 48 h, and examined directly under a microscope (b). Encysting cells overexpressing EGFP and AcSir2 at 24, 48 and 72 h after treatment with 0.5% sarkosyl for 30 min and staining with 0.4% trypan blue, with the number of mature cysts counted under a microscope (c). d Effects of salermide on the encystation of A. castellanii trophozoites. A. castellanii trophozoites were transferred into encystation medium containing 100 μM of salermide and incubated for 72 h. The mature cysts were counted under a microscope, followed by treatment with sarkosyl. DMSO was used as a solvent control. The percentage of mature cysts is calculated as the mean number of cells remaining after sarkosyl treatment compared with the total number of cells. The data represent the mean ± standard deviation (SD) of three separate experiments. **P < 0.01, ***P < 0.001 and ****P < 0.0001