CRACR2a cortical puncta are distinct from previously characterized endosomal and Golgi-derived vesicles. (A) CRACR2a cortical puncta (indicated by yellow arrowheads) do not colocalize with Rab5, Rab11, Rab22, Arf6, or clathrin-coated pits (indicated by white arrowheads). Scale bar: 3 µm. (B) Quantifications of the number of CRACR2a puncta observed in activated Jurkat cells under different treatments. DynK44A, coexpression of the K44A dominant-negative mutant of dynamin 1; n = 15–48 ROIs from at least 20 cells collected during three independent experiments. Error bar indicates SEM. ****, P < 0.0001, one-way ANOVA with Dunnett’s multiple comparisons test comparing each sample with mock (DMSO) treated sample.