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. 2018 Jan 15;32(2):96–111. doi: 10.1101/gad.309583.117

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© 2018 Li et al.; Published by Cold Spring Harbor Laboratory Press

This article, published in Genes & Development, is available under a Creative Commons License (Attribution 4.0 International), as described at http://creativecommons.org/licenses/by/4.0/.

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Figure 1. — Generation of an inducible EBF1 expression system in Ebf1^−/− pre-pro-B cells. (A) Schematic presentation of a 4-OHT-inducible retroviral EBF1 expression cassette. The retroviral loxP-Stop-loxP-Ebf1 construct contains a dsRed or tailless mCD8a reporter gene (R) and stop codon (red circle labeled with X) cassette that is flanked by loxP sites (orange triangles) and followed by an Ebf1 cDNA (green box). (LTR) Long terminal repeat. (B) Sorted pre-pro-B cells from the fetal livers of Ebf1^−/− RERT^Cre mice were transduced with loxP-Stop-loxP-Ebf1 retrovirus. EBF1 expression was induced by the addition of 2 µM 4-OHT, and cells were analyzed at the 24- and 72-h time points and at the CD19⁺ pro-B-cell stage. (C) Immunoblot analysis to detect the expression of transcription factors before and after EBF1 induction. (D) Flow cytometric analysis of intracellular EBF1 expression and the B-cell surface marker CD19 in Ebf1^−/− RERT^Cre::LoxP-Stop-LoxP-Ebf1 pre-pro-B cells before and after 4-OHT treatment.