Successes of populist radical right parties have impacted on the general political climate in Europe, and have resulted in the de-legitimation of several types of civil society organisations (CSOs) at all levels of governance, including at the EU level. This has resulted in diminished access to EU institutions, funding difficulties and lack of advocacy opportunities; civil society groups report a global reduction in a range of policy initiatives that have traditionally made use of their advocacy and campaigning skills. This lack of initiatives in turn translates into a diminished ‘space’ for contestation, which impacts specifically on those CSOs seeking to critique or support the EU, or articulate alternative visions of Europe. Associations particularly affected comprise those working on human rights, anti-discrimination groups focussing on gender and LGBT-rights, anti-racist and migrants’ advocacy groups. With reference to EU-level groups working on these issues, this paper examines their reactions to the emerging political climate and responses by the EU institutional framework within which they operate. The methodology consists of a frame analysis of a sample of position papers of civil society organisations, and a frame analysis of 20 in-depth interviews with EU-level civil society personnel and European Commission civil servants who interact with them. The paper argues that civil society reactions against populist parties should best be framed as a conflict between movements and counter-movements, which are proposing two opposed ideational constructs. With reference to the literature on discursive institutionalism, the paper documents the reasons that have hindered the formation of a cohesive response by several civil society groups. It explores their emerging opportunities and the strategies adopted to confront the new political climate, which vary across policy sectors and include partial de-politicisation of their advocacy efforts, institutionalisation and open conflict.