Skip to main content
. 2019 Jun 13;11(6):822. doi: 10.3390/cancers11060822

Figure 2.

Figure 2

CLL cells cultured in vitro are not directly affected by treatment with recombinant GRN. Data points in plots depict means of 2–5 technical replicates. The number of data points thus indicates biological replicates (n). Wilcoxon matched-pairs signed rank tests of samples with or without GRN treatment, all p values > 0.05. (a) Percentage of viable CLL cells in the indicated culturing conditions and treated with the indicated concentrations of GRN after 2 days (in low cell density (n = 1) or with 50 units/mL IL4 (n = 2)), 4 days (with 2 µM CpG oligodeoxynucleotides and 100 U/mL IL2 (n = 3)), or 9 days (in high cell density, n = 1) of culture was assessed by Annexin V/7AAD apoptosis assay via flow cytometry. (b) CLL cell viability was assessed by CellTiter-Glo assay after 2 days of incubation with the indicated concentrations of GRN (intermediate cell density, no additional factors added). RLU: relative luminescence unit. n = 6 biological replicates. (c) The fraction of proliferating CLL cells represented by EdU+ cells was assessed by Click-iT assay after treatment with 2 μM CpG and 100 U/mL IL2. n = 5 biological replicates. (d) CLL cell activation was measured by CD86 surface levels via flow cytometry after treatment with 100 ng/mL CD40 mega-ligand (CD40ML). 50 U/mL IL4 was added in all conditions. nMFI: normalized median fluorescence intensity (MFI CD86−MFI IgG isotype control). n = 6 biological replicates.