Extended Data Figure 3. Analysis of Vav-iCre/ROSA26-RARα403 mice.
a, E15.5 intestines from WT, Vav-iCre RarHet and RarHom mice were analysed by flow cytometry. Representative analysis of six independent experiments is shown. b, E15.5 regions of cervical, brachial and inguinal LN from WT, Vav-iCre RarHet and RarHom mice were analysed by flow cytometry. Representative analysis of two independent experiments. c, LTin cell percentage and LTi4/ILC4neg cell ratios are shown in E15.5 LNs; n=4. d, E15.5 foetal livers from WT and Vav-iCre RarHet mice were analysed by flow cytometry. Results show number of CD45+Lin− (n=4) and CD45−Lin−IL7Rα+α4β7+ progenitors (n=3). e, Percentage of CD45+CD3−CD11c−IL7Rα+RORγt+CD4− (RORγt+CD4−) and CD45+CD3−CD11c−IL7Rα+RORγt+CD4+ (RORγt+CD4+) cells determined by flow cytometry in RARHet and WT littermate controls in E15.5 guts and LNs. f, Frequencies of RORγt+CD4− and RORγt+CD4+ cells in mice described in (e) WT n=9; RarHet n=3. g, ILC4neg cells were purified from E15.5 WT intestines by flow cytometry and cultured for 6 days. LTi4 cells raised in vitro were purified by flow cytometry and quantitative RT-PCR analysis performed. Results show Log2 fold increase in comparison to their cultured ILC4neg cell counterparts. Results were normalised to Hprt1 and Gapdh. h. Left: E15.5 embryos were whole mount stained for CD4 (red) and imaged by confocal microscopy. Cervical (Cer) LNs are shown. Right: Cervical LN dimensions are shown. WT n=5; RarHet n=7; RarHom n=6. Scale bar: 50μm. Two tailed t-test p values are indicated. *P<0.05; **P<0.01; ***P<0.001. n.s., not significant.