Supplementary Figure 7: CLEC-2–Fc induces elongation and impairs spreading in a way similar to a genetic deletion of PDPN.

(a) Graph depicting the morphology index for untreated or CLEC-2-Fc-treated wild-type, Pdpn^−/−, and Δcyto FRCs. (b) The cell area of FRCs spreading on collagen in the presence or absence of CLEC-2-Fc. Data are representative of 3 independent experiments (mean±s.d., n>50 cells per experiment). (c) Graph representing the morphology index of wild-type FRCs that were pre-treated with cytochalasin D prior to CLEC-2-Fc treatment. (d,e) Representative images of YAP staining in wild-type (d) and Δcyto (e) FRCs that were untreated or treated with CLEC-2-Fc for 24 h. (f,g) Quantification of immunoblots for p-ERM (f) and p-MLC (g) levels in wild-type FRCs treated with CLEC-2-Fc for the indicated times. (h-k) Relative levels of inflammatory markers in LNs from mice treated with an isotype control or CLEC-2-Fc for 1 h. (l) Total cellularity from LNs of mice injected i.v. with an isotype control or CLEC-2-Fc for 1 h. (m) Morphology index for wild-type and Pdpn^−/− LECs treated with an isotype control or CLEC-2-Fc for 24 h. (n) Representative images of HEVs in LNs from isotype- or CLEC-2-Fc-treated mice. Red blood cells are stained with Ter119 and can be seen leaving HEVs in CLEC-2-Fc-treated LNs. NS, not significant; ^*P<0.01; ^**P<0.001; ^***P<0.0001 (Mann Whitney test).