Since the 1980s, the level of activism of regions in European Union policy-making has greatly increased, leading to the emergence of claims that regional governments can and do bypass national government in European negotiations. However, two decades after the emergence of the concept, the debate about the ability of regions to engage successfully in this process of continuous negotiation and to represent their interests on the European stage is ongoing. Due to the scarcity of research looking at regional interest representation in concrete cases of policy-making, it has been difficult to establish to what extent and under which circumstances regions do rely on unmediated channels of interest representation on the European level. This article examines these questions through the activities of seven legislative regions during two negotiations of European Directives, as legislative regions have a wider choice of channels of interest representation. Overall, extensive use of unmediated access in regulatory policy-making is rare and can best be explained with reference to domestic conflict and the level of influence of a region in domestic European policy-making. Differences in the size of a region also influence the ability of a region to represent its interests in the coordination of the national position and at the European level.