Fig. 4

The IL10-JAK-STAT3 positive feedback loop contributes to DLBCL motility. a, b Representative results of western blots of indicated proteins in whole-cell lysates (WCL), IL-6 and IL-10 in conditioned media (CM) from HT-S1 and HT-L1 sublines. Quantification of IL-6 and IL-10 expression. n = 3. Data represent mean ± S.D. c, d OCI-Ly3 cells were grown in collagen gels and treated with IL-6 or IL-10. Representative western blots and pull down assay for showing the level of indicated proteins. Quantification of RhoA activity. n = 3. Data represent mean ± S.D. e Quantification of movement speeds of OCI-Ly3 cells treated with IL-6 or IL-10 (n = 30). f Quantification of movement speeds of OCI-Ly3 cells treated with IgG, IL-6, or IL-10 neutralizing antibody (n = 30). g GSEA for showing the association between the signature of amoeboid movement and gene expression array from healthy peripheral blood mononuclear cells stimulated with IL-10 or control media (GSE43700). h GSEA for showing the association between the signature of amoeboid movement and gene expression array from monocyte-derived macrophages stimulated with IL-6 or control media (GSE8515). i HT cells expressing WT STAT3 or a control vector (puro) were injected into the spleen of SCID/beige mice. Mice bioluminescent signals on indicated days were detected (n = 8; other 5 mice in Supplementary Fig. 4i). j Quantification of disseminated numbers in i and Supplementary Fig. 4i. k-l Quantification of bioluminescent intensity of i and Supplementary Fig. 4i. from the primary sites (yellow dotted lines circled area) and the disseminated sites (red dotted lines circled area). NS, not significant. m Quantification of the circulating tumor cells (CTCs) collected from the mice described in panel i. n = 8. n HT cells expressing WT STAT3 or a control vector (puro) were intraperitoneal injected into SCID/beige mice for 9 weeks. The dot plot shows the serum concentrations of IL-10. n = 6. o Distribution of the H score for IL-10 expression from 73 DLBCL patients. p Representative results of immunohistochemical staining for IL-10. Scale bar = 20 μm.*, P < 0.05; **, P < 0.005 by Student’s t-test; Kolmogorov–Smirnov tests for GSEA. See also Supplementary Fig. 4