Supplementary Figure 6: Analysis of iT cells in Tet-Hoxb5 pro-pre-B recipients

(a) Schematic diagram of targeting strategy using Tet-Hoxb5 transgenic model for conditional expression of Hoxb5. The indicated expression cassette Tet-Hoxb5-BFP was inserted into ROSA26 locus. The BFP reporting Hoxb5 can be induced by Doxycycline (Dox). (b) Schematic strategy of B to T conversion by transient expression of Hoxb5 using Tet-Hoxb5 model. (c-d) Fow cytometry analysis of the iT cells gated from Ter119^-Mac1^-CD19^-CD45.2⁺ in the PB and spleen of Tet-Hoxb5 pro-pre-B cell recipients maintained on Dox-water (1 mg/mL) for four weeks (c) and recipients maintained on conditions of Dox withdrawal 2 weeks prior to analysis (d). Wild-type CD45.2 C57BL/6 mice were used as negative control. Recipient mice were analysed four weeks post-transplantation. (e) Q-PCR of ectopic Hoxb5 in BFP^- and BFP⁺ thymic CD3⁺ T cells from Tet-Hoxb5-NOD-SCID mice. Wild-type CD45.2 C57BL/6 CD3⁺ thymic T cells were used as negative control (n = 6 biological replicates). Data are representative of three independent experiments (c, d).