Fig. 2

Inhibition of TGF-β-mediated cancer cell migration and macrophage polarization by Mϕ-SDNP. (A) Viability of 4T1 cells after treatment with PLNP, SDNP, MϕNP, and Mϕ-SDNP as determined using MTT assay. (B) Flow cytometric analysis for cellular uptake post Cy5.5-loaded MϕNP treatment. ***p < 0.001. Statical significance was calculated with Student’s t-test (n = 3). (C) Representative confocal fluorescence microscope images. Nuclei were stained with DAPI. Scale bars = 15 μm. PLGA nanoparticles (PLNP), Integrin α4-blocked macrophage membrane-coated nanoparticle (Ib-MϕNP), and macrophage membrane-coated nanoparticles (MϕNP) were treated. D - E) SD-208-loaded MϕNP (Mϕ-SDNP) significantly reduced TGF-β-mediated cancer cell invasion. (D) Wound healing assay. 10 μg/ml of each nanoparticle was applied, and TGF-β was treated at a concentration of 20ng/ml. Scale bars = 100 μm. (E) Transendothelial migration assay. The HUVEC cell monolayer migrated cancer cells were stained with crystal violet. The relatively small dot represents migrated cancer cell. Scale bars = 100 μm. (F) EMT marker gene expression analysis. The mRNA expression was normalized by mouse GAPDH expression. (G) Schematic representation of the BMDM isolation and M2-type differentiation process. H-I) Relative mRNA expression analysis of (H) M2 macrophage marker and (I) M1 macrophage marker. The mRNA expression was normalized by mouse GAPDH expression. ***p < 0.001. Statistical significance was calculated with Student’s t-test