Flow cytometry analysis of bone marrow erythroid cells.

<p>Bone marrow cell suspensions were prepared from dissected tibiae and femora from mice fed control (0.6% Pi) or low phosphorus diet (0.02% Pi), as described in Coe <i>et al</i> 2014. For immunostaining, cells were first incubated with CD16/32 antibody to block mouse Fc receptor and reduce non-specific binding. Erythroid lineage was assessed using Ter119-APC / CD71-PE markers combined with the forward scatter (FSC) properties, as described in Asari S <i>et al</i> 2005 Exp Hematol, and Koulnis M <i>et al</i> 2011 J Vis Exp. Labeled cells were then analyzed by flow cytometry. Appropriate isotype controls were kept for each set. Forward and side scatter patterns were gated excluding the debris. A total of 20,000 events were collected and analyzed using FlowJo software.</p> <p>(PDF)</p>