Fig. 1.

BMP4 favours in vitro proliferation and pro-inflammatory cytokine production. (a) pSMAD1/5 was detected by western blot in Jurkat cells cultured in the presence of BMP4 (500 ng/ml) for 30 min; the phosphorylated form of the protein decreased or disappeared when noggin, DM or DMH1 was added to the culture medium in the presence of BMP4. (b) Splenocytes from naïve mice were cultured in the presence of BMP4 (10 μg/ml) or a combination of BMP4 and DM (1 μM) or DMH1 (40 μM). BMP4 induced splenocyte proliferation, while proliferation was abrogated in the presence of DM and DMH1, which are BMP signaling inhibitors. The dotted line represents the proliferation threshold. Data correspond to three independent experiments. (c) Cytokines were quantified in the supernatants of splenocytes cultured with BMP4, BMP4+DM, or BMP4+DMH1. The dotted line indicates the lower limit of quantification for each cytokine. Data are represented as the mean ± standard error of the mean (SEM). Data were analyzed with mixed models to account for clustering by experiment and repeated measures. All conditions were compared with the BMP4-treated cultures using Dunnett’s adjustment. BMP4 = bone morphogenetic protein 4; CM = culture medium; DM = dorsomorphin; DMH1 = dorsomorphin homologue 1