Fig. 6.

Podoplanin, CD44 and CD9 are required for FRCs to respond to CLEC-2⁺ DCs. (A) Immunofluorescence of 3D cultures of indicated FRC cell lines with LPS-stimulated bone marrow-derived DCs (CD45⁺; magenta). Maximum Z-stack projections of representative images from n=2 biological replicates are shown. Scale bars: 20 μm. (B) Morphology index (perimeter²/4π area) of DCs in interaction with an FRC. Dots represent single DCs. n=40-72 DCs collated from two biological replicates. Error bars represent median with interquartile range. *P=0.0149; ns, not significant (Kruskal–Wallis test with Dunn's multiple comparisons). The y-axis has a log₁₀ scale. (C) Immunofluorescence of 3D cultures of FRC cell lines without (upper row) or with (middle and bottom rows) LPS-stimulated bone marrow-derived DCs (magenta). Maximum Z-stack projections of representative images from n=2 biological replicates are shown. Scale bars: 50 μm. (D,E) F-actin intensity (mean grey value of phalloidin–TRITC staining; D), and morphology index (perimeter²/4π area; E) of indicated FRCs alone (open circles) or in interaction with a DC (closed circles). Dots represent single FRCs. n=36–104 FRCs collated from two biological replicates. Error bars represent median with interquartile range. ****P<0.0001; ns, not significant (two-way ANOVA with Tukey's multiple comparisons).