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This method was used to improve our understanding of the ordered assembly of the Saccharomyces cerevisiae spindle pole body (SP, a ~1 giga-Dalton heteromeric protein complex formed from 18 structural components present in multiple copies. We propose that SRM-BiFC is a powerful tool for examination of direct interactions between protein complex subunits at sub-diffraction resolution in live cells.Non-covalent binding of cholesterol to the transmembrane region of proteins affect their functionalities, but methods to prove such an intera