Fig. 6: TGF-β impairs granulysin expression and secretion in population B.

a Flow cytometry analysis of surface expression of CD127, CD117 and CD18 in population B (CD127+CD94+CD117−). Cells were cultured for 7 days on OP9 cells, which were excluded from the analysis by gating for live CD45+ cells as depicted in supplemental Fig. 4a, in the presence of IL-2 (10 U/ml) and IL-15 (50 ng/ml) with or without TGF-β (50 ng/ml). Dot plots depict mean percentage ±SEM of CD117+ cells, CD127+ cells (both N = 5) and CD18+ cells (N = 2). Each dot represents one donor. b Flow cytometry analysis of intracellular expression of cytotoxic molecules granulysin, granzyme B and perforin in population B after culturing for 7 days on OP9neo, which were excluded from the analysis by gating for live CD45+ cells, in the presence of IL-2 (10 U/ml) and IL-15 (50 ng/ml) with or without TGF-β (50 ng/ml). Dot plots depict the mean relative MFI ± SEM of Granulysin (N = 6), granzyme B (N = 5) and perforin (N = 3) upon addition of TGF-β compared to the condition without TGF-β. Each dot represents one donor. c Concentration of granulysin in the supernatant of cells from population B cultured on OP9neo for 7 days in the presence of IL-2 (10U/ml) and IL-15 (50 ng/ml) with or without TGF-β (50 ng/ml) measured by ELISA. Dot plot depicts the mean concentration ±SEM, each dot represents one donor (N = 3). All Data were tested for normal distribution using the Shapiro-Wilk test and if normally distributed tested with a paired t-test. If not normally distributed (perforin) data were tested with a Wilcoxon matched-paired rank test. All p values are two-sided.