Fig. 4.

Satb2 and Etv5 downstream of aPKCλ regulates B cell differentiation arrest. a Heat map of the B-cell differentiation network in WT and DKO leukemic B-cell progenitors whole transcriptome. b–d Q-RT-PCR analyses of genes [Pax5 (b), Ebf1 (c), and Cdkn2a (d)] that are involved in B-cell differentiation program in leukemic B-cell progenitors derived from WT, aPKCζ^−/−, aPKCλ^Δ/Δ, and DKO chimeric mice. e Representative example of western blots showing the expression of chromatin modifier, Satb2, and actin in WT, aPKCζ^−/−, aPKCλ^Δ/Δ and DKO leukemic B-cell progenitors. f Q-RT-PCR analysis of SATB2 in human BM B progenitors (CD34⁺/CD19⁺) from normal individuals (n = 2) and Ph⁺ B-ALL pediatric patients (n = 3). g Chromatin immunoprecipitation using α-Satb2 antibody and Q-PCR analyses of genes involved B-cell differentiation program Pax5, Ebf1, and Cdkn2a in p210-BCR-ABL expressing leukemic B-cell progenitors. Leukemic B-cell progenitors show enhanced enrichment of Pax5, Ebf1, and Cdkn2a loci. We found no enrichment of the housekeeping genes Gapdh and Actb. h Representative example of western blots showing expression of Etv5 in WT, aPKCζ^−/−, aPKCλ^Δ/Δ, and DKO leukemic B-cell progenitors. i Chromatin immunoprecipitation using Etv5 antibody and Q-PCR analyses of Stab2 promoter enrichment in leukemic B-cell progenitors. We found no enrichment of the housekeeping genes Gapdh and Actb. j–k Chromatin immunoprecipitation using α-H3K27me3 (j) and α-H3K4me3 (k) antibodies and Q-PCR analyses of the enrichment of Pax5, Ebf1 and Cdkn2a genes in WT and aPKC deficient B-cell  progenitors. DKO leukemic B-cell progenitors show enhanced activation of genes involved in B-cell differentiation in comparison to their WT counterparts. Cont: Control. Data are presented as mean ± SD of a minimum of two independent experiments. *p < 0.05; **p < 0.01; ***p < 0.001, t-test