Fig. 2: EZH2 inhibitor-treated BC cells activate TAMs to a M2 phenotype. | Cell Death & Disease

Fig. 2: EZH2 inhibitor-treated BC cells activate TAMs to a M2 phenotype.

From: Regulation of CCL2 by EZH2 affects tumor-associated macrophages polarization and infiltration in breast cancer

Fig. 2

A Scheme of the workflow. 4T1 cells labeled with GFP, a green tracking dye were transiently transfected with siEZH2 or treated with EPZ-6438, and seeded in a 3D culture qualified 96 well spheroid formation plate for spheroid formation. After 3-6 days, spheroids were formed and 100 µl of invasion matrix containing BMDM cells polarized by IL-4 and IL-13 treatment and labeled with mCherry, a red tracking dye were added. The cell number ratio of 4T1 and BMDM was close to 4:1. Representative photographs of infiltrating macrophages into the 4T1 microsphere (magnification, ×20) were shown B and relative mCherry luminescence signals were quantified C. Scale bar represents 100 μm. D Scheme of the workflow. BC cells were transiently transfected with siEZH2 or treated with EPZ-6438. CM was used as a chemoattractant for macrophages in a transwell migration assay. E The relative transcript levels of the M2-related genes in the CM-treated BMDM were determined by RT-qPCR. F The migration rates of BMDM-derived M2 macrophages with or without the co-culture of 4T1 CM were determined by transwell assay. G The expression of M2 macrophage markers in co-cultured BMDMs were tested by western blot. The graphs were shown as means ± SD, n = 3 independent experiments; *p < 0.05; **p < 0.01; ***p < 0.001, compared with control, p values were obtained using a two-tailed Student’s t test.

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