Eukaryotic translation initiation factor 5A and its posttranslational modifications play an important role in proliferation and potentially in differentiation of the human enteric protozoan parasite Entamoeba histolytica
Fig 9
Steady state transcript levels of EieIF5A isoforms during excystation.
(A) Kinetics of excystation. Cysts were incubated at the indicated times, cysts and trophozoites were harvested and the excystation percentage was determined. The excystation efficiency was determined by counting trophozoites and cysts in a haemocytometer and experiments were performed in duplicate. Data shown are the means ± standard deviations of two independent experiments. (B) Semi-quantitative RT-PCR analysis of EieIF5A1, EieI5FA2, EieIF5A1 and EiActin genes during excystation. cDNA from 4 time points during excystation was subjected to 30 cycles of PCR using specific primers for eIF5A1, eI5FA2, eIF5A3, and actin genes. Actin gene served as a control. PCR from samples without reverse transcription served as controls to exclude the possibility of genomic DNA contamination. The densitometric quantification of the bands, shown in the right graph, was performed by Image J software, and the expression level of EieIF5A1, EieIF5A2, and EieIF5A3 and EiActin was expressed in arbitrary units.