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Blood, Vol. 113, Issue 24, 6128-6137, June 11, 2009 This Article Abstract Full Text Services Email this article to a friend Alert me to new issues of the journal Rights and Permissions Citing Articles Citing Articles via CrossRef Cytoskeletal protein 4.1R negatively regulates T-cell activation by inhibiting the phosphorylation of LAT Blood Kang et al. 113: 6128 Supplemental materials for: Kang et al Files in this Data Supplement: Table S1. Phenotyping of major lymphoid organs (PDF, 17.3 KB) Figure S1. 4.1R is recruited to the interface between T-cell– and antigen-presenting cell (JPG, 40.9 KB) - (A) Primary T-cell blasts from 5c.c7 transgenic mice were stained with anti-4.1R exon 13 polyclonal antibody, followed by anti-rabbit Alexa-488–conjugated secondary antibody, and analyzed by confocal microscopy. Note that protein 4.1R is evenly distributed around the cell membrane. (B) Primary T-cell blasts stimulated by peptide-loaded CH27 cells were stained as described above. Note that protein 4.1R becomes enriched at the interface between the T cell and the APC cell. Scale Bar, 2 µm. Images shown in both panel A and panel B are collage of images from four different fields. Figure S2. Expression of 4.1R in thymocytes and enhanced signaling in 4.1R−∕− thymocytes (JPG, 36.5 KB) - (A) themocytes (1 × 106 cells) from 4.1R+∕+ or 4.1R−∕− mice were subjected to immunoblot analysis with polyclonal rabbit antibodies against 4.1R exon 13. The position of ~ 135 kDa and ~ 80 kDa 4.1R is indicated. (B) 4.1R+∕+ or 4.1R−∕− thymocytes were stimulated with anti-CD3ε (5 µg/ml) for varying time periods, and indicated proteins were immunoblotted for the level of expression and for the extent of phosphorylation. (C)The enhanced phosphorylation of both LAT and ERK in 4.1R−∕− thymocytes is significant. Figure S3. SDS-PADE of recombinant proteins used in the present study (JPG, 21 KB) - 1 µg of GST or His fusion protein was analyzed by 10% SDS-PAGE and stained with GelCode Blue. This picture is a collage from two gels. Figure S4. Interaction of cytoplasmic domain of LAT with Lobe C of 4.1R as assessed by surface plasmon resonance assay (JPG, 49 KB) - GST-tagged Lobe C of 4.1R was immobilized onto CM5 sensor chip. His-tagged cytoplasmic domain of LAT at different concentrations was injected at 20 µl/min over the surface in a BIAcore 3000 instrument. The figure shows dose-response curves of LAT binding to the immobilized Lobe C. This Article Abstract Full Text Services Email this article to a friend Alert me to new issues of the journal Rights and Permissions Citing Articles Citing Articles via CrossRef

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