Extended Data Fig. 8. Phenotypic and functional characterization of SYK^S544 mice.

(a) Immunoblot analysis of SYK and pSYK protein levels in ankle of 3 months old mice (wt: n=3; SYK^S544Y: n=3). (b) Hematoxylin and eosin stain of tail tissue sections from 3 months old wild-type and SYK^S544 mice showing bone erosion and immune cell infiltration (wt: n=3; SYK^S544Y: n=3). (c) Hyper-phosphorylation of SYK in intestinal tissue from SYK^S544 mice compared to wild-type mice (wt: n=3; SYK^S544Y: n=3). (d) Western blot analysis of wild-type and SYK^S544 bone marrow derived dendritic cells SYK phosphorylation (Y519/520), total SYK expression, JNK1 (T183/Y185) phosphorylation, total JNK1 expression, ERK1 phosphorylation (T203/Y205) and total ERK1 expression treated or not treated with R406 SYK inhibitor (2 μM, R406 was added 30 minutes prior to LPS (200 ng/mL) stimulation and samples collected after 24 hours) (wt: n=3; SYK^S544Y: n=3). (e) Analysis of IgG and IgM in serum from wild-type and SYK^S544Y mice by ELISA at indicated age (n=3; Unpaired t-test). (f) RT–qPCR analysis of Il17f (wt: n=3; SYK^S544Y: n=6), Csf2 (wt: n=3; SYK^S544Y: n=9), Ifng (wt: n=3; SYK^S544Y: n=3), Il17a (wt: n=6; SYK^S544Y: n=6) and Il4 (wt: n=3; SYK^S544Y: n=3) expression in blood at the age of 3 months. Unpaired t-test. (g) Serum cytokine concentrations in wild-type and SYK^S544Y mice at 3 months of age measured by ELISA (n=4; unpaired t-test). Violin plots indicate quartiles and median.