Supplementary Figure 4: Ablation of FRCs disrupts the organization of splenic white pulp and humoral responses of the marginal zone.

(a) Confocal analysis of spleen sections from Ccl19-Cre x Rosa26-EYFP mice stained for GFP, B220 and CD4. Scale bars 200 μm (left) and 50 μm (right) (n=3 mice from two separate experiments). (b-e) Spleens from Ccl19-Cre x Rosa26-EYFP-iDTR mice were processed into single cell suspensions and EYFP⁺cells were enumerated using flow cytometry. (mean+/-sd, n=3 mice). (f,g) Weight and total cell numbers for spleens of Ccl19-Cre x iDTR mice are indicated. Data are representative of at least two independent experiments (mean+/-sd, n=3 mice/group per experiment). (h) Confocal analysis of splenic white pulp from Ccl19-Cre x iDTR animals 24 h following DTxn administration. Scale bars 200 μm (n=3 mice from two independent experiments). (i) B cells in the spleen were enumerated using flow cytometric analysis (CD45⁺B220⁺CD3^–). Data are representative of at least three independent experiments (mean+/-sd, n=3 mice/group per experiment). (j) Flow cytometric analysis of lymph nodes and spleen in DTR⁺Cre^– and DTR⁺Cre⁺ animals. Fold decrease in FRC numbers at indicated time points is shown. (k) Marginal zone humoral responses were assessed by determining antigen-specific IgM production via ELISA following TNP-ficoll immunization. ^NS non significant, *p<0.05, **p<0.01

Source data