Apc conjugated rat anti mouse cd44

Bone marrow (BM) was collected and processed as described previously [21 (link)]. Cells were stained with APC-conjugated rat-anti-mouse CD44 (BD Pharmingen, Heidelberg, Germany) and PE-conjugated rat-anti-mouse TER119 (BD Pharmingen, Heidelberg, Germany) in 2% FBS/PBS for 30 min at RT protected from light. Analysis was performed using the BD FACSDiva™ software on a FACSCalibur™ (Becton Dickinson, Heidelberg, Germany). Unstained cells were used as negative controls. Mean fluorescence intensity (FLI) from 20.000 cell counts was used as a measure of protein surface expression of Ter119 and CD44 [22 (link)–24 (link)].

Bone marrow (BM) was collected and processed as described previously [26 (link)]. BM cells were stained with APC-conjugated rat anti-mouse CD44 and PE-conjugated TER119 rat anti-mouse (BD Pharmingen, Heidelberg, Germany) in PBS/2%FBS for 30min at RT. Analysis was performed using the BD FACSDiva^™ software on a FACSCalibur^™ (Becton Dickinson, Heidelberg, Germany). Unstained cells were used as background control.
Ter119+ events were considered as erythroid precursors which were further discriminated by cells size (forward scatter) and CD44 expression. CD 44 expression declines as erythroid precursors reach later stages of differentiation [27 (link)].