Fig. 4.

Identification of ternary complexes with BiFC–FRET approach. a Schematic representation of BiFC-based FRET approach adopted here detecting the ternary complexes between the medial Golgi enzymes, the NSTs and the mutual interactions between the MGATs and NSTs. b BiFC–FRET analyses of the depicted enzyme and transporter constructs. Cells were triple transfected with the constructs, fixed 24 h later before quantification of the FRET signal using the Operetta High Content Imaging System. As negative controls (light grey columns) we utilized different combinations, each of which contained one non-interacting construct (B4GALT1, AE2). Statistical significance of the changes was calculated using one-way ANOVA with Tukey’s Post Hoc test (± SD, n = 3, ***p < 0.001)