Fig 6. Determination of cytoplasmic extract dilution for hemagglutination and yeast agglutination inhibition assays.

Cytosolic extract containing CV-IIL or RS-IIL_A22S was serially diluted in working buffer, and each sample from the dilution line was mixed with 5% RBC_0⁺ or yeast cell suspension in a 1: 1 ratio. The mixture was incubated at room temperature for 5 or 10 minutes, respectively, mixed again, applied to a glass slide and observed under a Levenhuk microscope. Pictures were taken with a DEM135 camera (Levenhuk). 128× diluted and 2× diluted cytoplasmic extract was chosen for the CV-IIL hemagglutination and yeast agglutination inhibition assay, respectively. 32× diluted and undiluted cytoplasmic extract were chosen for the RS-IIL_A22S hemagglutination and yeast agglutination inhibition assay, respectively. All negative control experiments did not exhibit any visible agglutination. The positive control was done using purified CV-IIL.