Fig. 5.

Forced expression of Etv5 and Satb2 rescue B cell differentiation arrest. a Schematic diagram of Etv5 and Satb2 ectopic expression in Lin⁻cKit⁺Sca1⁺ (LSK) cells derived from Scl/p210 primary chimeric mice and co-culture on a OP9 mouse stromal cells monolayer and/or in vivo transplantation into secondary recipients. b Q-RT PCR showing expression of Etv5 in mock/Etv5 transduced B progenitors. c Q-RT PCR analyses showing rescue of Satb2 expression in Etv5 transduced B progenitors. d FACS quantification of EGFP⁺ (mock/Etv5/Satb2 expressing) B-cell progenitors derived from OP9 stromal monolayer co-culture of mock/Etv5 transduced LSK cells. e, f Representative spleen images (e) and spleen weight (f) of mock/Satb2 transduced and transplanted secondary recipient mice. g Satb2 ectopic expression enhances the CFU- preB content. h, i FACS quantification of EGFP⁺ B-cell progenitors content in the bone marrow (BM) (h) and spleen (i) of secondary recipient mice transplanted with mock/Satb2 transduced LSK cells. j, k Q-RT PCR analyses of Satb2 (j) and Etv5 (k) in mock or Satb2 expressing B progenitors. l–n Q-RT PCR analyses of expression of B-cell differentiation genes Pax5 (l), Ebf1 (m) and Cdkn2a (n) in mock or Satb2 expressing B-cell progenitors. o Chromatin immunoprecipitation using α-H3K27me3 antibody and Q-PCR analyses of the enrichment of Pax5, Ebf1, and Cdkn2a genes in mock/Satb2 transduced WT/DKO leukemic B-cell progenitors. p Chromatin immunoprecipitation using α-H3K4me3 Ab and Q-PCR analyses of the enrichment of Pax5, Ebf1, and Cdkn2a genes in mock/Satb2 transduced WT/DKO leukemic B-cell progenitors Data are presented as mean ± SD of a minimum of three independent experiments.*p < 0.05; **p < 0.01; ***p < 0.001, t-test