Interplay between the EMT transcription factors ZEB1 and ZEB2 regulates hematopoietic stem and progenitor cell differentiation and hematopoietic lineage fidelity
Fig 4
Double deletion of Zeb1 and Zeb2 causes PB cytopenia and severe differentiation defects in HSPCs.
(A) Schematic of experiments (left panel) used to study the effect of tamoxifen-inducible deletion of Zeb1, Zeb2, or both after donor BM reconstitution (CD45.2+) of lethally irradiated recipients (CD45.1+). Schematic of loxP flanked (floxed-Fl) conditional Zeb1 and Zeb2 alleles before tamoxifen induced Cre-mediated deletion and recombined delta Zeb1 and Zeb2 alleles after recombination (right panel). (B) HCT analysis of BM 10 days after tamoxifen treatment showing decreased HGB (top left) in Zeb1Δ/Δ, Zeb2Δ/Δ, and Zeb1/2Δ/Δ DKO settings. There was as well decreased RBC (top middle), PLT (top right), and GRA (bottom left). LYM (bottom middle) in the BM were decreased only in the Zeb1/2Δ/Δ DKO. (C) Flow cytometric analysis of HSPCs in the BM 10 days after tamoxifen treatment showing increased numbers of LSK cells and well as LT-HSCs (LTS- lin−cKit+Sca1+CD34−Cd125−) in Zeb1/2Δ/Δ DKO settings. (D) Representative flow cytometry analysis for hematopoietic stem and progenitor populations showing increased numbers of LSK and LT-HSCs in Zeb1/2Δ/Δ DKO settings. Bars in panels represent mean ± SD, n = 5 per group; *p < 0.05; **p < 0.01; ****p < 0.0001, Dunnett multiple comparisons test. Raw data behind graphs are included in C of S1 Data. BM, bone marrow; DKO, double knockout; GRA, granulocyte; HCT, hematocrit; HGB, hemoglobin; HSPC, hematopoietic stem and progenitor cell; LSK, Lin−Sca1+cKit+; LT-HSC, long-term HSC; LTS, long-term HSC; LYM, lymphocyte; MPP, multipotent progenitor; PB, peripheral blood; PLT, platelet; RBC, red blood cell; STS, short-term HSC.