Impact of BCOR on erythroid and granulocytic differentiation. (A) Evolution of BCOR gene expression during normal granulocytic and erythroid differentiation. BCOR messenger RNA level as measured by reverse transcriptase quantitative PCR (RT-qPCR) is shown as normalized relative quantities (NRQs) ± SEM to B2M and ACT. (B) shRNA-mediated BCOR silencing in normal human CD34⁺ progenitors (day 7). shSCR as control. Western blot to BCOR; actin is used as loading control. (C) Proliferation of short hairpin BCOR (shBCOR) or shSCR human CD34⁺ progenitors was driven by IL-3, SCF, FLT-3L, and TPO for 18 days. Results are representative of 4 independent experiments. (D) Clonogenic progenitor growth. shBCOR or shSCR CD34⁺ progenitors were amplified in liquid culture for 6 days and seeded in methylcellulose. Colony numbers ± SEM. (E) (Left) Immunophenotypic quantification of CD71, GPA, Band3, and CD49d expression in shBCOR and shSCR cells at day 18 of erythroid differentiation. Biparametric histograms are shown and percentages are indicated. Results representative of 3 independent experiments. (Right) GATA1 and HBB gene expression according to expression of shBCOR or shSCR by RT-qPCR. Results expressed as NRQs to UBC and ACT ± SEM represent 2 independent experiments in duplicates. (F) (Left) Immunophenotypic quantification of CD33 and CD11b. (Right) SPI1 and MPO gene expression by RT-qPCR.