Skip to main content
. Author manuscript; available in PMC: 2016 Jul 1.
Published in final edited form as: Exp Parasitol. 2015 Apr 15;154:33–42. doi: 10.1016/j.exppara.2015.04.004

Figure 2. Overexpression of EhROM1-SA and downregulation of EhROM1 in E. histolytica trophozoites result in reduced cytotoxicity and hemolytic activity.

Figure 2

(A and B) Cytotoxicity was measured by placing a total of 0.5×105 trophozoites on a confluent CHO cell monolayer for 2h at 37°C, followed by fixation, staining with 0.1% methylene blue, dye extraction, and spectrophotometric determinations at 650 nm. Overexpression of EhROM1-SA as well as downregulation of EhROM1 resulted in parasites with decreased ability to destroy CHO cell monolayer. *p-value <0.001 control versus EhROM1-SA; *p-value <0.039 control versus T-EhROM1-s. (C and D) Hemolytic activity was measured by incubating human erythrocytes with trophozoites in 100:1 ratio for 1h and released hemoglobin quantified at the absorbance of 405 nm. Overexpression of EhROM1-SA and downregulation of EhROM1 resulted in parasites with decreased hemolytic activity. *p-value <0.0005 control versus EhROM1-SA; *p- value <0.018 control versus T-EhROM1-s. The results represent the mean and standard deviations of three to five independent experiments and are expressed as the percentage of the control strain destruction level or hemolytic activity. The control cell line is one that is overexpressing (A and C) or silencing (B and D) the luciferase gene. The G418 drug concentration in (A and C) is at 12μg/ml and in (B and D) is at 6μg/ml.