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. 2017 Jun 23;7:4156. doi: 10.1038/s41598-017-04537-4

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© The Author(s) 2017

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.

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Loss of Gnai2 inhibits early thymocyte development and causes SP mature thymocytes to accumulate. (A) Representative flow cytometry of thymocytes from indicated mice examining CD4 versus CD8 expression. (B) Representative flow cytometry of thymocytes from indicated mice gated on CD4 SP thymocytes for their expression of CD62L and CD69. (C) Representative flow cytometry of thymocytes from indicated mice gated on DN thymocytes for their expression of CD44 and CD25. (D) Growth and differentiation of FACS sorted DN1(Lin⁻CD25⁻ CD44⁺CD117⁺) thymocytes purified from the indicated mice and cultured on OP9-DL1 cells in the presence of IL-7 (1 ng/ml) for 28 days. The numbers of total and DN thymocytes recovered at Day 28 from each of the genotypes is shown to the right. (E) Growth of FACS sorted DN3 thymocytes from the indicated mice cultured on OP9-DL1 cells in the presence of IL-7 (1 ng/ml) for 7 days. Experiments were performed a minimum of 3 times. **p < 0.005 (Student’s t-test). (F) Representative flow cytometry plots from part D comparing WT and Gnai2 ^−/− DN1 thymocytes cultured for various durations on OP9-DL1 cells.