Using multiple representations and argumentation are two fundamental processes in science. With the advancements of information communication technologies, these two processes are blended more so than ever before. However, little is known about how these two processes interact with each other in student learning. Hence, we conducted a design-based study in order to distill the relationship between these two processes. Specifically, we designed a learning unit on nuclear energy and implemented it with a group of preservice middle school teachers. The participants used a web-based knowledge organization platform that incorporated three representational modes: textual, concept map, and pictorial. The participants organized their knowledge on nuclear energy by searching, sorting, clustering information through the use of these representational modes and argued about the nuclear energy issue. We found that the use of multiple representations and argumentation interacted with each other in a complex way. Based on our findings, we argue that the complexity can be unfolded in two aspects: (a) the use of multiple representations mediates argumentation in different forms and for different purposes; (b) the type of argumentation that leads to refinement of the use of multiple representations is often non-mediated and drawn from personal experience.