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. Author manuscript; available in PMC: 2018 May 16.
Published in final edited form as: Immunity. 2017 May 16;46(5):818–834.e4. doi: 10.1016/j.immuni.2017.04.022

Figure 4. Lymphoid Organs and Tissues Derived from Tcf3fl/flTcf12fl/flIl7rCre Mice show Increased ILC2 Cellularity.

Figure 4

(A) Cell numbers of ILC2s in the BM, lymph nodes (LNs), and spleen derived from 4-week-old littermate control and Tcf3fl/flTcf12fl/flIl7rCre mice (n=5, cumulative from 4 independent experiments). (B) Flow cytometric analysis of Gata3 expression in ILC2s in thymi, BM, LNs and spleens. Red indicates isotype control whereas blue refers to Gata3 expression (representative from two independent experiments). (C) Frequency and ILC2 numbers in lungs derived from 4-week-old littermate control and Tcf3fl/flTcf12fl/flIl7rCre mice are indicated (n=4, cumulative from two independent experiments). (D) Flow cytometric analysis of BrdU incorporation (left) gated on thymic ILC2s derived from 4-week-old Tcf3fl/flTcf12fl/flIl7rCre mice. Mice were injected intraperitoneally with 1mg of BrdU (blue) or PBS (red) twice (24 h and 2 h prior to harvesting). Flow cytometric analysis of Ki67 expression (right) gated on thymic ILC2s derived from 4-week-old Tcf3fl/flTcf12fl/flIl7rCre mice. IgG control (red), Ki67 (blue) (representative from two independent experiments). (E) Representative flow cytometric analysis of ST2 and ICOS expression gated on Lin−Thy1.2+CD25+IL-7R+Sca1+ thymocytes derived from adult E47−/−, Tcf12fl/flIl7rCre, and control mice. Numbers in plots indicate percentages ILC2s and cell numbers of ILC2s in thymi. (F) Total thymocyte number, frequency and cell numbers of ILC2s in the thymi derived from 4-week-old E47−/− and control mice are shown (n=4, from three independent experiments). (G) Frequencies and cell numbers of ILC precursors in thymi isolated from 4-week-old E47−/− and control mice are shown (n=3, from two independent experiments). (H) Gating strategy for thymic ILC2 cells in Rag2−/−, Tcf3fl/flTcf12fl/flIl7rCreRag2−/−, and Tcf3fl/flTcf12fl/flIl7rCre Rag2+/− mice. (I) Cell numbers of ILC2s, LTi-like cells, and ILC precursors in the thymi derived from Rag2−/− and Tcf3fl/flTcf12fl/flIl7rCreRag2−/− mice (n=3–5, from three independent experiments). Data represent the mean ± SD. *, P < 0.05, **, P <0.01, ***, P <0.001, ****, P <0.0001 (Student’s t test). See also Figure S3.