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. 2016 Feb 18;7(2):e2103. doi: 10.1038/cddis.2016.23

Figure 3.

Figure 3

mRNA and protein expression levels of A1 in differentiating neutrophils. (a) relative mRNA expression levels of A1 during differentiation. Wt Hoxb8 progenitor cells were induced to undergo differentiation by estrogen withdrawal and monitored daily up to day 4. Cells were harvested at the indicated time points, followed by extraction of total RNA and reverse transcription into cDNA. Quantitative real-time RT-PCR was performed on a Light Cycler 2 instrument. Shown are A1 mRNA expression levels relative to β-actin expression normalized to progenitors (day 0). Data represent mean/S.E.M. of a total of 3–4 independent experiments. *P≤0.05. (b) Protein expression levels during differentiation of Hoxb8 neutrophils. Progenitor cells were induced to undergo differentiation by estrogen withdrawal either in the presence of SCF alone or with additional stimulation with G-CSF. Cells were harvested at the indicated time points and extracted by lysis and boiling in Laemmli buffer. Whole-cell lysates were separated by SDS-PAGE, transferred onto nitrocellulose membrane and probed for A1 and Mcl-1. GAPDH served as a loading control. Shown is one representative experiment out of three (SCF alone) or two (SCF+G-CSF)