Thymic phenotype of Themis−/− Shp1 cKO mice is similar to that of Themis−/− mice. Thymic phenotype of Themis−/− Shp1 cKO mice is similar to that of Themis−/−

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Thymic phenotype of Themis−/− Shp1 cKO mice is similar to that of Themis−/− mice. Thymic phenotype of Themis−/− Shp1 cKO mice is similar to that of Themis−/− mice. (A) Western blot showing amounts of Themis and Shp1 in thymocytes of Themis+/+, Themis−/−, Shp1fl/fl CD4-Cre+ (Shp1 cKO), and Themis−/− Shp1 cKO mice. Total Erk is used as a loading control. Shp1 was quantified by normalizing to total Erk. (B) Representative flow cytometry showing different subpopulations of thymus (Top) and thymus maturation based on CD69 and TCR-β (Bottom) in Themis+/+, Shp1 cKO, Themis−/−, and Themis−/− Shp1 cKO thymocytes. (C) Graphs showing enumeration of total number (*106) of CD4 SP, CD8 SP, TCRhiCD69hi, and TCRhiCD69lo populations in the thymus in Themis+/+, Shp1 cKO, Themis−/−, and Themis−/− Shp1 cKO mice. Data are representative of five experiments. There were three to four mice per genotype of the same age and sex in each experiment. Unpaired t test with mean ± SD. **P < 0.01; ****P < 0.0001. ns, not significant. (D) Western blot showing levels of p-Erk on stimulation with Kb tetramers in Shp inhibitor-treated and untreated thymocytes from OT-I β2m−/−Themis+/+ and OT-I β2m−/−Themis−/− mice. Raw values of proteins were calculated using LI-COR Odyssey software with the help of the background subtraction method. Monika Mehta et al. PNAS 2018;115:48:E11331-E11340 ©2018 by National Academy of Sciences