Thymic lymphoproliferation in FU-Lcktg/Lckind mice. (A) Dot plots showing distribution of T-cell subsets in thymi from 12-week-old Lcktg/Lck−/−/Lckind and FU-Lcktg/Lck−/−/Lckind mice defined by surface CD4 and CD8 expression and FACS analysis. Values within dot plots represent proportions of cells with DP and CD8 SP gates. Dot plots represent staining from 1 of at least 5 mice of each genotype. FACS histograms showing cell size (B) as assessed by analysis of forward scatter (FSC) and surface expression of αβTCR (C), CD5 (D), and CD24 (E) on gated CD4+CD8+ DP cells and CD8+ SP cells from thymi of Lcktg/Lck−/−/Lckind and FU-Lcktg/Lck−/−/Lckind mice. Filled histograms represent Lcktg/Lck−/−/Lckind mice; black line overlays represent FU-Lcktg/Lck−/−/Lckind mice. Data represent 1 of at least 5 mice of each genotype. (F) FACS dot plots showing distribution of T-cell subsets in thymi from 12-week-old F5 Rag1−/−/Lck−/−/Lckind, FU-Lcktg/Rag1−/−/Lck−/−/Lckind, and FU-Lcktg/F5/Rag1−/−/Lck−/−/Lckind mice. Values represent the proportions of cells within DP and CD8 SP gates as indicated. Data represent 1 of at least 7 mice of each genotype. Cell size (G) and surface expression of αβTCR (H) on DP and CD8 SP cells. Filled histograms represent F5 Rag1−/−/Lck−/−/Lckind mice; black line overlays represent FU-Lcktg/F5/Rag1−/−/Lck−/−/Lckind mice. (I) Comparison of cell surface and intracellular αβTCR FACS staining of thymocytes from Lck−/−/Lckind and FU-Lcktg/Lck−/−/Lckind mice and CD19+ splenocytes from FU-Lcktg/Lck−/−/Lckind mice. For intracellular staining, cells were permeabilized in 0.5% saponin before staining with αβTCR mAb. Filled histograms represent levels of surface αβTCR; black line overlays represent levels of staining after cell permeabilization. (J) PCR analysis of TCR Dβ2.1-Jβ2.7 rearrangements. The figure represents a 1.2% agarose gel of PCR products from B6 ear,1 B6 thymus,2 and 3 individual 10-week-old FU-Lcktg/Lck−/−/Lckind thymi.3-5 The 100-bp markers are also shown (M).
