Fig. 5.

Aged Gal-3 KO mice presented signs of autoimmunity. a ANA IgG titres determined using Hep-2 slides in sera from 8-month-old WT and Gal-3 KO mice. Magnification: ×100 (top) and ×600 (bottom) (WT n = 9; Gal-3 KO n = 13). Scale bar, 200 μm (upper panels) and 20 μm (lower panels). b dsDNA IgG determined using Crithidia luciliae slides. Magnification: ×1200 (WT n = 9; Gal-3 KO n = 13). Scale bar, 10 μm. c Pie charts summarizing the distribution of serum samples exhibiting the different Hep-2 patterns: Nucleolar; nuclear homogeneous; nuclear fine speckled; nuclear coarse speckled; and non-HEp-2 Reactive. The numbers in the centers of the graphs denote the number of samples analysed per group. Animals with ANA titres higher than 1/40 were scored as positive (WT n = 9; Gal-3 KO n = 13). d IgM, IgG1, IgG2c, and IgG3 autoantibodies levels specific for dsDNA, histone and Sm/RNP determined by ELISA (WT n = 5; Gal-3 KO n = 7). e Kidney sections stained with H&E (left) and the nephritis score (right). Magnification: ×100 and ×400 (WT n = 10; Gal-3 KO n = 12). Scale bar (from left to right), 100, 20, 100, and 20 μm. f Immunofluorescence of kidney sections stained with anti-B220 (green), anti-CD45 (red) and anti-CD4 (blue). Magnification: ×200 (n = 4 for both strains). Scale bar, 100 μm. g, h Urea (left) and creatinine (right) concentrations in sera (g) and protein levels in 24-h urine samples (h) from 8-month-old WT and Gal-3 KO mice (WT n = 4; Gal-3 KO n = 7). i IFN-γ and IL-17 concentrations in sera from 8-month-old WT and Gal-3 KO mice (n = 5 for both strains). Data are the mean ± s.e.m. *p < 0.05, **p < 0.005, ***p < 0.001. ns not significant (unpaired Student’s t-test). All experiments were performed in 8-month-old WT (green) and Gal-3 KO (blue) mice. a, c, e Data are representative of four independent experiments. b, d Data are representative of two independent experiments. f–i Data are representative of three independent experiments