Protocol for assessing protein expression levels and localization during fission yeast cytokinesis

The fission yeast Schizosaccharomyces pombe is a genetically tractable model that shares mechanisms for cytokinesis with higher eukaryotes. Here, we present a protocol for analyzing fission yeast cytokinesis proteins. We include steps for gene targeting by homologous recombination, functionality tests by genetic interactions, and imaging by high-spatiotemporal resolution microscopy. Next, we describe procedures for image analyses using Fiji, protein interactions by co-immunoprecipitation, and measuring protein levels by Western blotting. These techniques are transferable to other subcellular locations and model organisms. For complete details on the use and execution of this protocol, please refer to Ye et al.¹.