Fig. 4: EZH2 increases osteolytic cytokines expression in myeloma cells.

a A published GEO dataset (GSE103567) analysis shows the expression of cytokines in RPMI8226 cell treated with or without GSK343 (4 μM). b, c The expression of CCL5, IL16, CSF1 and DKK1 in RPMI8226 (shCtrl, shEZH2) or MM.1S (Vec, EZH2) cells (n = 3 biological replicates). d–g ELISA analysis shows the secretion of CCL5, IL16, CSF1 and DKK1 in RPMI8226 (shCtrl, shEZH2) or MM.1S (Vec, EZH2) (n = 3 biological replicates). Correlation coefficient of the levels of CCL5 (h), IL16 (i), CSF1 (j) or DKK1 (k) and levels of EZH2 (n = 30). The cells used here are from the same source as those in Fig. 1b. The correlation was evaluated using Pearson coefficient with two-tailed p value. r, correlation coefficient. l The numbers of multinuclear TRAP⁺ cells among precursors of osteoclasts cultured with MM.1S cells (Vec or EZH2) in the presence of blocking antibodies against CCL5, IL16, CSF1 or DKK1 (1 μg/ml) (n = 3 biological replicates). m The Alizarin red S staining of MSCs cultured in osteoblast medium with MM.1S cells (Vec or EZH2) cells in the presence of antibodies against CCL5, IL16, CSF1 or DKK1 (1 μg/ml) (n = 3 biological replicates). n The numbers of multinuclear TRAP⁺ cells among precursors of osteoclasts cultured with RPMI8226 cells (shCtrl or shEZH2) in the presence of CCL5 (20 ng/ml), IL16 (2 ng/ml), CSF1 (10 ng/ml) or DKK1 (20 ng/ml) (n = 3 biological replicates). o The Alizarin red S staining of MSCs cultured in osteoblast medium with RPMI8226 cells (shCtrl or shEZH2) cells in the presence of CCL5 (20 ng/ml), IL16 (2 ng/ml), CSF1 (10 ng/ml) or DKK1 (20 ng/ml) (n = 3 biological replicates). Data are means ± SD. b–g: p values were determined by unpaired two-tailed t test; l–o: p values were determined using one-way ANOVA with Tukey’s test. Source data are provided as a Source data file.