Cd71 surface markers

We used neutrophilic-differentiated NB4 cells (NB4^PMN) to study the effect of hyperresistinemia on neutrophil function in a standard and reproducible way. Neutrophilic differentiation was achieved by previously described methods [10 (link)]. Briefly, NB4 cells were treated with 1 μM all-trans retinoic acid (ATRA) for 6 days in culture media. Successful neutrophilic differentiation was assessed using CD11b, CD35, and CD71 surface markers (BD Biosciences) with flow cytometry. For some experiments, NB4^PMN were incubated for 1 h in human control serum (Atlanta Biologicals Flowery Branch, GA, USA) spiked with different concentrations of recombinant human resistin (R&D Systems, Minneapolis, MN, USA) at 20, 50, 100 ng/mL to replicate normal circulating vs. septic resistin concentrations.

THP-1 monocytes were differentiated into macrophages (THP-1^mac) by incubation with 100 nM phorbol 12-myristate 13-acetate (PMA) for 72 h [17 (link)]. Differentiated cells were incubated in fresh RPMI media for 24 h before initiation of experimental studies. Prior to experimentation, successful macrophage differentiation was confirmed by expression of CD14, CD16, and CD71 surface markers (BD Biosciences) with flow cytometry.