Fig. 3

Establishment of pre-induced thymic epithelial cell (pre-iTEC) line and induction of differentiation into thymic epithelial cells (iTEC). A. Experimental scheme: Genetic circuits were introduced into iPSCs to create pre-induced thymic epithelial cells (pre-iTEC). Induction into induced thymic epithelial cells (iTEC) was achieved by administering 1 µg/mL doxycycline for 7 days. B. Representative images of the created pre-iTEC1 and PGP1 cells. The RFP signal indicates the expression of the mScarlet gene. C. RT-qPCR comparison of the introduced genes rtTA, BSR, and mScarlet between groups (n = 3). D. RT-qPCR comparison of undifferentiated marker genes OCT4, NANOG, and DNMT3B in PGP1 cells and pre-iTEC1 (n = 3). E. RT-qPCR comparison of FOXN1 gene expression in pre-iTEC1 and pre-iTEC1 treated with doxycycline groups (n = 3). F. Morphological changes in pre-iTEC1 upon doxycycline administration. G. RT-qPCR comparison of undifferentiated markers in pre-iTEC1 with and without doxycycline treatment (n = 3). H. Comparison of TEC markers DLL4 and ENPEP (Ly51) in pre-iTEC1 upon doxycycline treatment (n = 3). I. Representative image of immunofluorescence staining for DLL4 in pre-iTEC1 treated with doxycycline. P-values were calculated using a two-tailed unpaired t-test. The data are presented as mean ± SEM. For detailed data, statistical analyses, and exact p-values, see source data file