Figure 1
Thymocyte development and lymphopoiesis are impaired at critical checkpoints in the absence of 2A-DUB/Mysm1. (a) Mysm1 mRNA expression of indicated thymocyte subsets FACS sorted from 4- to 6-week-old C57BL/6 mice relative to GAPDH (left two bar graphs). Induction of Mysm1 mRNA in thymocytes and splenocytes stimulated with αCD3 (1 μg/ml) for indicated time periods and thymocytes 4 h after exposure to etoposide (eto, 10 μM) or γ-irraditation (γ, 4 Gy) relative to GAPDH (right bar graphs). In all qPCR analyses, bar graphs represent mean expression of a least three mice and two individual experiments ±S.D. (b–e) Flow cytometric analysis of total (CD19, Mac1, Gr1, Ter119, NK1.1, and Epcam1-negative) or lineage (CD4, CD8, CD3, CD19, Mac1, Gr1, Ter119, NK1.1, and γδTCR)-negative (Linneg) thymocytes or Linneg BM cells from 4- to 6-week-old male Mysm1−/− (KO) mice in comparison with age-matched WT littermates (dot plots from representative experiments are shown, n>8). Total cell numbers (shown in bar graphs next to the FACS profiles) of indicated subsets represent averages from at least six mice and three independent experiments ±S.D. (f) Dot plots and histograms of CD25 and Foxp3 expression of indicated thymocyte subsets and correlation with Foxp3 mRNA expression of total Mysm1+/+ (WT) or Mysm1−/− (KO) thymocytes. FACS data show representative examples of at least three separate experiments with two WT (grey lines) and two KO (grey dotted lines) thymi relative to isotype control Ab (grey area). Bar graphs show means of three experiments ±S.D.
