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. 2007 Dec 3;179(5):923–934. doi: 10.1083/jcb.200708200

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Copyright © 2007, The Rockefeller University Press

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Figure 2. — IP₃R1 phosphoY353 forms a spatially restricted signaling microdomain with TCR. (A, top) Calnexin staining of ER in stimulated T cells in cell spreading assay. Jurkat T cells were incubated on CD3-coated coverslips for 5 min and stained for calnexin (green), TCR/CD3 (red), and total phosphotyrosine (blue). (bottom) 3D rendering of image slice shown on top. The 3D rendering reveals global distribution of ER upon T cell activation. In the cell spreading assay, ∼70% of cells appeared to be stimulated (formed a central TCR cluster). Of that population, almost all the cells showed colocalization of Y353 with the TCR. (B, top) Staining of IP₃R1 phosphoY353 in stimulated T cells. Jurkat T cells were incubated on CD3-coated coverslips for 5 min and stained for IP₃R1 phosphoY353 (green), TCR/CD3 (red), and total phosphotyrosine (blue). (bottom) 3D rendering of image slice shown on top. The 3D rendering reveals localization of IP₃R1 phosphoY353 to a discrete site colocalizing with TCR and phosphotyrosine with localization potentially mediated by the extrusion of the ER membrane. Bars, 5 μm.