<i>Cebpa</i> knockdown reduces murine granulopoiesis relative to monopoiesis and increases immature blasts in liquid culture.

<p><b>A</b>) Transduced, puromycin selected marrow cells were placed in liquid culture with IL-3, IL-6, and SCF on D0 and viable cell counts were enumerated on days 0, 2, 3, 4, and 7. Representative data is shown. <b>B</b>) Cell cycle analysis was conducted using PI staining on D0 or D3. <b>C</b>) FACS analysis for Mac-1 and Gr-1 on D2. Arrows indicate immature, Mac-1⁻Gr-1⁻ cells (top). FACS analysis of Mac-1⁻Gr-1⁻ cells stained for Sca-1 and c-Kit (middle). FACS analysis of D2 cells for FcγR and CD34, gating on Lin⁻Sca-1⁻c-Kit⁺ progenitors (bottom). Representative data are shown. <b>D</b>) Total cellular RNAs collected on day 3 (D3) after lentiviral transduction with the pLKO.1 vector or with <i>Cebpa</i> shRNAs B9 or B11, puromycin selection, and transfer to IL-3, IL-6, and SCF were subjected to quantitative RT-PCR for indicated mRNAs (n = 3).