Figure 2.

Characterization of spleens from WT and Ats1-KO mice. (a) Graph representing the spleen index (as indicated in the Methods section) of healthy WT (n = 6) and Ats1-KO (n = 5) mice to assess splenomegaly. Inset below include representative images of spleens from WT and Ats1-KO mice (white scale bar = 1 cm). (b) Graph representing flow cytometry data, as percentage of positive cells of the following populations: CD45R⁺, CD3⁺, CD11b⁺, GR1⁺, CD11b⁺/GR1⁺, and F4/80⁺, found in spleens of healthy WT (n = 5) and Ats1-KO (n = 5) mice. (c) Graph representing the mRNA fold change expression of Cd3g, Cd4, Il6, Il10 and Il12a genes in spleens of healthy WT (n = 5) and Ats1-KO (n = 5) mice, being all values relative to WT. (d) Representative images of spleen sections from WT and Ats1-KO mice, showing VCAN and VKIN (red) (left and right column, respectively), and CD31 (green) immunofluorescence staining. Images correspond to a 63x magnification (white scale bar = 20 μm). (e) Western blot analysis with anti-DPEAAE antibody for ADAMTS-cleaved versican in spleen protein extracts from WT and Ats1-KO mice. Blue arrow indicates a predicted versican (V0) fragment (around 220 kDa). Bottom panel includes actin staining (grey arrow). Full-length blots are presented in Supplementary Figure 4. (f) Graph representing the absolute Ct value for mRNA expression of Adamts1, 4, 5, 9, 15, and 20 genes in spleens of healthy WT (n = 5) and Ats1-KO (n = 4) mice (values higher than 37–38 are considered very low or absent). For all the graphs, results are shown as the median with s.e.m. and statistical significance (*p < 0.05; **p < 0.01; ***p < 0.001; ****p < 0.0001).