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. 2013 Jun 25;8(6):e65896. doi: 10.1371/journal.pone.0065896

Figure 3. “Re-educated” macrophages affected 4T1 cell proliferation, cell cycle, cell apoptosis, migration and invasion.

Figure 3

(A) Trypan blue staining demonstrated mortality of 4T1 cells incubated with PBS, co-cultured with E-RAW264.7, E-RAW264.7 treated with plain NPs (NPs-E-RAW264.7) or E-RAW264.7 treated with 18 µM HC-NPs (RE-E-RAW264.7) for 12 hours respectively. (B) (C) Flow cytometry detected cell cycle and cell apoptosis of 4T1 cells incubated with PBS, co-cultured with E-RAW264.7, NPs-E-RAW264.7, RE-E-RAW264.7 for 12 hours respectively. X Parameter, Annexin-v (Log); Y Parameter, PI (Log). Data are representative of 3 separate experiments. (D) (E) Migration and invasion of 4T1 cells significantly decreased after co-cultured with RE-E-RAW264.7. Transwell migration and invasion assays, cells were stained with 0.1% crystal violet solution and analyzed with ImageJ. Original magnification: × 200. (F) The “re-educated” macrophages affected p-STAT3, STAT3, Caspase-3, Bcl-2, PARP, MMP-9, MMP-2 and VEGF protein expression of 4T1 cells after RE-E-RAW264.7/4T1 co-culture.