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. 2016 Sep 30;197(9):3520–3530. doi: 10.4049/jimmunol.1600959

FIGURE 1.

FIGURE 1.

Generation of a cd4-1 reporter line. (A) Schematic illustrating the construction of the transgene. A BAC containing the 5′ region of the cd4-1 gene was selected and modified by recombination to contain Tol2 transposable elements with the kanamycin resistance gene (dark blue box) to allow selection. A cassette containing the mCherry coding sequence and the spectinomycin resistance gene (SpecR) flanked by cd4-1 homology arms (HA1 and HA2) was then recombined into the cd4-1 start codon (located in exon 3). (B) The Tg(cd4-1:mCherry) reporter is expressed in the thymus by 5 dpf and colocalizes with the pan–T cell reporter lck:GFP. (C–E) High magnification view of the thymus at 5 dpf revealing CD4-1+ (mCherry+ GFP+) and CD4-1− (GFP+ only) T cells. (F) Reporter expression in the thymus of adult fish at 2 mo postfertilization in the casper mutant background. Anterior is to the left. Scale bar, 10 μm. Denoted features are eye (ey), otic vesicle (ot), thymus (th).