Fig 1: The recovery culturing downregulates expression of CD16 and CD56 on NK cells. (a) Representative immunoblot of CD16A (FcγRIIIa) in the whole cell lysates of NK cells isolated from unrecovered (NR) PBMCs or from PBMCs after recovery cultivation for different time points (4 h, 24 h); (b) Quantitation of three independent CD16A immunoblots; (c) Flow cytometry analysis of surface CD16 (FcγRIII) on live NK cells isolated from fresh (neon green), thawed cryopreserved (NR, orange) PBMCs, or from PBMCs after recovery cultivation for different time points 4 h (red) and 24 h (blue). Data in this figure were analyzed with GraphPad Prism using one-way ANOVA mixed—effects statistical analysis based on matched rows and recovery time as repeated measure with Tukey’s post hoc test. p = 0.0279; (* p < 0.05). (d) Quantitation of replicated flow cytometry results shown in (c) p = 0.0708; (e) Flow cytometry analysis of NK cells isolated from unrecovered (NR) PBMCs or from PBMCs after recovery cultivation for different time points (4 h, 24 h); (f) Quantitation of replicated flow cytometry results shown in (e) p = 0.0037 (** p < 0.01) (time factor) within CD56dim/CD16dim, p = 0.0040 (** p < 0.01) (time factor) within CD56bright/CD16bright. Fresh NK cells were prepared from a different donor.