This data set contains the raw and analysed data used for the research article: SNX9-induced membrane tubulation regulates CD28 cluster stability and signalling. This includes raw fluorescence microscopy, electron tomography and flow cytometry files, as well as MS excel and Graphpad PRISM analysed data. Please use the "Organisation of the data" pdf file for more detailed information about the content of the folders. Abstract of the article: T cell activation requires engagement of a cognate antigen by the T cell receptor (TCR) and the co-stimulatory signal of CD28. TCR and CD28 aggregate into clusters at the plasma membrane of activated T cells. While the role of TCR clustering in T cell activation has been extensively investigated, little is known about how CD28 clustering contributes to CD28 signalling. Here we report that upon CD28 triggering, the BAR-domain protein sorting nexin 9 (SNX9) is recruited to CD28 clusters at the immunological synapse. Using three-dimensional correlative light and electron microscopy, we show that SNX9 generates membrane tubulation out of CD28 clusters. Our data further reveal that SNX9 regulates CD28 cluster stability as well as CD28 phosphorylation and the resulting production of the cytokine IL-2. In summary, our work suggests a model in which SNX9-mediated tubulation generates a membrane environment that promotes CD28 triggering and downstream signalling events.