Fig. 7: Synovial fibroblasts isolated from inflamed and non-inflamed joints have distinct sialylation signatures.

Synovial fibroblasts were isolated from healthy mice (naive) and from CIA arthritic mice, where joints were separated into non-inflamed (low scores, LS) and very inflamed (high scores, HS) prior to cell isolation and expansion. a N-glycans were isolated and permethylated for to MALDI-TOF MS analysis and relative glycan expression was quantified as in Fig. 3. Two clusters were identified as distinctive of severe inflammation, for which glycan structures are shown. Percentage of sialylated and non-sialylated glycans are represented. Cell lines from a were stimulated with TNF (b) and IL-17 (c) [10 ng/ml, 12 h]. Supernatants were collected and IL-6 levels were quantified by ELISA. Each dot represents an independent experiment analyzed in triplicate. Data are presented as mean ± SEM, n = 3 (IL-17), and n = 4 (TNF) independent experiments. Statistical significance was evaluated by ordinary one-way ANOVA and Tukey’s test for multiple comparations. ***p < 0.001, **p < 0.01, *p < 0.05. Actual p-values: a <0.0001, b <0.0001, c 0.0265, d 0.0008. ^#p < 0.05 compared with naive non-treated, actual p-values: e 0.003, f 0.00028, g 0.0393, h 0.0158. d Cell lines were stimulated with galectin-3 [1 ng/ml, 12 h] and IL-6 was measured in culture supernatants by ELISA. Data are presented as mean ± SEM, n = 6 independent experiments. One-tail unpaired t-test was used for statistics, *p < 0.05. Actual p-values: i 0.0476, j 0.0243.