Figure 5
From: Salt inducible kinases 2 and 3 are required for thymic T cell development
SIK2-/-/SIK3fl/fl/Vav-iCre+/- mice show abnormal T cell development. (A) Thymi were isolated from wild type (WT) and SIK2-/-/SIK3fl/fl/Vav-iCre+/- (KO) mice and analysed by flow cytometry. Data show the number of Thy1.2+ve cells in the thymus, representative CD4/CD8 flow cytometry plots of Thy1.2+ve cells in the thymus and the numbers of DN, DP, CD4 SP and CD8 SP cells. (B–D) Splenocytes were isolated from wild type (WT) and SIK2-/-/SIK3fl/fl/Vav-iCre+/- (KO) mice and analysed by flow cytometry following lysis of red blood cells. Single cell suspensions were analysed for expression of CD3, CD4, CD8, CD44 and CD62L. Total cell numbers and numbers of CD3+ve T cells in the spleen, along with representative FACS plots are shown in (B). Absolute numbers of CD4 and CD8 T cells along with representative CD4/CD8 plots of CD3+ve T cells are shown in (C). The expression of CD44 and CD62L was also examined in both CD3+ve/CD4+ve and CD3+ve/CD8+ve T cells and the data shows representative flow cytometry plots and the percentage of CD44+ve/CD62L-ve and CD44-ve/CD62L+ve cells (D). Graphs show mean with symbols representing measurements from individual mice. Differences in cell number between wild type and knockout mice were analysed by Student’s t-test (B and Thy1+ve cells in A) or RM two way ANOVA with Sidak’s post hoc testing in all other graphs. p < 0.05 is indicated by *, < 0.01 by ** and < 0.001 by ***. For the thymus 11 WT and 12 knockout mice were analysed while for the spleens 8 mice were examined per genotype.
