Figure 4
From: A complex of Neuroplastin and Plasma Membrane Ca2+ ATPase controls T cell activation
PMCA1 and PMCA4 are Neuroplastin-binding partners. (A) Table showing Neuroplastin interaction with PMCA1 and 4 in thymocytes revealed by MS of Neuroplastin-immunoprecipitates (IP) from wt thymocyte solubilizates. No unique peptides were detected for PMCA2 or 3. (B) Verification of the Neuroplastin-PMCA complex by Western blot detection of PMCA1 and 4 in Neuroplastin-IP. Asterisks mark immunoglobulin light and heavy chain bands. Nptn −/− control samples confirmed specificity. (C) Immunofluorescent double labeling of Neuroplastin (magenta) and PMCA1 (green) and co-staining for DNA (DAPI, blue) on conjugates formed between pOva-loaded B cells and OT-II transgenic wt (upper panel) or Nptn −/− (lower panel) CD4 T cell blasts, scanned at 63x magnification and shown as 3D reconstructions from confocal stacks. Merged images reveal striking co-localization (white) of both proteins at the synaptic interface between the elongated wt T cell (T) and the weakly labeled B cell (B). Note that reduced PMCA1 in Nptn −/− T cells still enriches at the immune synapse.
