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. 2020 Sep 7;27(12):3226–3242. doi: 10.1038/s41418-020-00615-9

Fig. 6. EZH2-R342 methylation promotes breast cancer cell invasion and metastasis.

Fig. 6

a, b The mRNA and protein expression of EMT markers were detected by qRT-PCR and western blot in MCF7-vector/Flag-EZH2-WT/Flag-EZH2-R342K cells. c The mRNA expression of EMT inducers in MCF7-vector/Flag-EZH2-WT/Flag-EZH2- R342K cells. d Western blots analysis of MMP9 and MMP2 expression in MCF7-vector/Flag-EZH2-WT/Flag-EZH2-R342K cells. e ChIP analyses on E-cadherin promoter were done using anti-EZH2 and anti-H3K27me3 antibodies in MCF7-shCtrl and MCF7-shPRMT1#1 cells. f–h Assessment of cells motility by wound-healing assays (f), invasion assays (g) and migration assays (h) in MCF7-vector/Flag-EZH2-WT/Flag-EZH2-R342K cells. i, j Representative bioluminescence images of lung metastasis (i) in mice injected with MM-231-luc-shEZH2-Vector/Flag-EZH2-WT/Flag-EZH2-R342K cells via tail veins, and the metastases were analyzed by measuring the photo flux (j), n = 9 for each group. k–n Lung metastatic nodules were examined macroscopically (k and l) or detected by H&E staining after nude mice were sacrificed (m and n). The white arrows denote the metastatic nodules. o, p Representative images of the IHC staining of meR342-EZH2 (o) or EZH2 (p) in lung metastasis sections. Data are represented as mean ± SEM of three or four independent experiments, and *p < 0.05, **p < 0.01, ***p < 0.001 (Student’s t test).