Fig. 5

Colocalization and binary interaction between GlNSF and the giardial α-SNAPs. a Colocalization of GlNSF with α-SNAP₁₇₂₂₄ or α-SNAP₁₀₈₅₆ in 48 h encysting trophozoites, and with α-SNAP₁₆₅₂₁ or α-SNAP₁₀₈₅₆ in trophozoites. Insets depict magnification of the ROI (marked with a white box). The scattergram in each row indicates the analysis of colocalization between the two fluorophores over the entire z-stack by considering the pixels within the entire area occupied by the particular cell. The values for Pearson correlation coefficient (Pr) and overlap coefficient (OC) are indicated inside the scattergrams. The intensity plots at the extreme right indicate changes in the intensity of the red and green fluorescence signals across the diagonal of the ROI depicted by a dotted white line. b The bar graph denotes the mean Pr and OC calculated from the z-stacks of six independent images. c PJ69-4A cells were transformed with various combinations of constructs expressing fusion proteins with either Gal4 DNA binding domain (BD) or its activation domain (AD). Expression of the BD or AD alone served as negative controls. Transformants were spotted on YCM plates lacking leucine and tryptophan (LT), or leucine, tryptophan and histidine with 2.5 mM 3-AT (LTH 3-AT), or leucine, tryptophan and adenine (LTA). d β-galactosidase activity of the indicate transformants was quantified. Statistical significance of the difference in interaction between any two interacting pairs is indicated (**P < 0.01, ***P < 0.001). Scale-bar: a, 5 μm