Fig. S1. EhCaBP6 binds Ca2+ in vitro. Ruthenium red staining: The indicated amounts of purified proteins were spotted on to a PVDF membrane in duplicate. These were then stained with ruthenium red dye as described under Experimental procedures.


Fig. S2. Sub-cellular localization of EhCaBP6 varies during the cell cycle of E. histolytica.

A. Localization of EhCaBP6 during the cell cycle of E. histolytica: Serum synchronized cells were fixed at different time points during the cell cycle of E. histolytica. This was followed by staining the fixed cells with anti-CaBP6 anti-CaBP6 primary and Alexa-488 conjugated anti-mouse secondary antibodies. DNA was visualized by staining with DAPI. The images were taken in LSM510Meta confocal microscope under 63× oil DIC objective.

B. mRNA levels of EhCaBP6 during different time points of E. histolytica cell cycle: RNA was isolated from serum synchronized E. histolytica cells. Semi-quantitative RT-PCT was done on the isolated RNA samples. The results have been displayed in a bar graph (Error bars, ± SD, n = 3).


Table S1. List of the proteins used to construct phylogenetic tree.


Table S2. Summary of macroscopic binding constants and Thermodynamic parameters obtained from the Isothermal calorimetry (ITC) studies of Ca2+-binding isotherm of EhCaBP6 at 25°C.

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