Figure 6.

Substrate stiffness regulates CD83 expression, but not T-cell activation capacity, of mDCs. (a,b,c) Cell surface expression of CD86 (a, n = 7 donors), CD83 (b, n = 7 donors) and MHC-II (n = 5 donors) in day 4 iDCs (gray bars) and day 2 mDCs (black bars) conditioned on PAA substrates of 2, 12 and 50 kPa, represented as geomean fluorescence intensity measured with flow cytometry. Maturation was induced by addition of IL-4, GM-CSF, IL-1β, TNF-α, IL-6 and PGE₂. Bars represent mean with SEM. Statistical significance among iDC/mDC conditions was tested with repeated measures ANOVA. (d) T-cell proliferation of allogeneic PBLs stimulated by day 2 mDCs (n = 2 donors, technical triplicates) conditioned on PAA substrates of 2, 12 or 50 kPa (at a ratio of 10:1), measured with ³H-Thymidine incorporation detected in counts per minute. Negative control contained no stimulus (medium only) and positive control contained anti CD3/CD28 T-cell activating beads at a ratio of 4:1. Bars represent mean with SEM. *p < 0.05; **p < 0.01.