Identification of a Novel Alternatively Spliced Form of Inflammatory Regulator SWAP-70-Like Adapter of T Cells

Figure 1

Identification of SLAT2, a novel SLAT splice variant. (a) CD4⁺ T cells from B6 mice were differentiated under Th1- or Th2-inducing conditions with spleen-derived APCs and anti-CD3 Ab for 5 days, and the cells were then restimulated with plate-bound anti-CD3 plus -CD28 mAbs. Cell lysates were prepared at the indicated days and analyzed by Western blotting using anti-SLAT Ab. Data are representative of three independent experiments. (b) Gel electrophoresis of RT-PCR products amplified from Th2 cells two days after the second round of stimulation using primers spanning the entire SLAT open reading frame. Arrows indicate the position of SLAT2 (lower) or a predictable full length SLAT (upper). (c) Expression of SLAT and SLAT2 proteins in differentiated Th2 cells and Jurkat-TAg cells transfected with pEF-SLAT or pEF-SLAT2. Differentiated Th2 cells from B6 mice were restimulated with plate-bound anti-CD3 plus -CD28 mAbs for 4 days. Cell lysates were analyzed by anti-SLAT or anti-Myc immunoblotting. Data are representative of three independent experiments. (d) Genomic structure of SLAT and the alternatively spliced isoform SLAT2. Primers used for PCR amplification are indicated with arrows. Exons and introns are shown as open boxes and lines, respectively. SLAT2 contains the DH-like domain but lacks the part of EF-hand and the entire ITAM-like sequence and PH domain.

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