Figure 3.

Aberrant T-lineage gene expression in progenitor B-cells deficient for Btg1 and Btg2. (A) Relative expression levels of B-cell transcription factors E2A (Tcf3), Foxo1, Ebf1 and Pax5 was determined using cDNA generated from B220⁺ bone marrow (BM) cells of wild type control (WT), Btg1^−/−, Btg2^−/− and Btg1^−/−; Btg2^−/−mice by qRT-PCR and normalized to the expression of the housekeeping gene TBP. (B) RNA was isolated from B220⁺CD43⁺ and B220⁺CD43⁻ cells derived from wild type control and Btg1^−/−; Btg2^−/− mice (n=4) and analyzed by microarray using the Illumina BeadArray platform. The data obtained were RMA normalized and relative gene expression differences of >1.6-fold was determined. KEGG pathway analysis was performed using the online Gene Set Enrichment Analysis (GSEA) tool and relevant pathways with highest P-value that are differentially expressed between WT and Btg1^−/−; Btg2^−/− cells are indicated. (C) Relative expression levels of Cd4, Ikzf2, Tcf7, Gata3 and Notch1 were determined on cDNA generated from B220⁺ BM cells of the different genotypes by real-time PCR and normalized to TBP expression. (A and C) Data represent the mean and SEM of three independent experiments containing cDNA derived from 2 different biological samples. *P<0.05, **P<0.01, ***P<0.001.