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. 2014 Apr 21;9(4):e95784. doi: 10.1371/journal.pone.0095784

Figure 4. Cebpa knockdown reduces murine granulopoiesis relative to monopoiesis and increases immature blasts in liquid culture.

Figure 4

A) 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) Cell cycle analysis was conducted using PI staining on D0 or D3. C) 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. D) Total cellular RNAs collected on day 3 (D3) after lentiviral transduction with the pLKO.1 vector or with Cebpa 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).