Figure 3.

Amigo2 induction in RA synoviocytes cocultured with immune cells remains stable even after immune cell removal. RA synoviocytes were cocultured with PBMC from healthy donors in the presence or not of PHA for 24 h. In the cocultures, PBMC were separated from synoviocytes by EDTA addition. Synoviocytes were cultured for 24 or 48 h more after PBMC removal. Amigo2 expression was assessed at different time-points by quantitative real-time PCR and was expressed as fold changes compared to RA synoviocytes cultured alone and exposed to vehicle (A). The production of IL-17A (B) and TNF-α (C) by the cocultures was quantified by ELISA. The production of these cytokines was measured in cocultures with no immune cell removal (cocultures) or with immune cell removal by EDTA (cocultures EDTA). Their production was not detectable in synoviocytes cultured alone. Data are the mean of at least three independent experiments ± SEM. ^#comparison with control situation, *comparison between synoviocytes (A) or PBMC (B) alone and cocultures, ^§comparison between cocultures with or without EDTA addition. *,#,§P ≤ 0.05, **,##,§§P ≤ 0.01, ***, ###P ≤ 0.001.