Supplementary Information Figures from Macrophage-Secreted TNFα and TGFβ1 Influence Migration Speed and Persistence of Cancer Cells in 3D Tissue Culture via Independent Pathways

Abstract

<p>This file contains figures in the supplementary information. These data complements data in the main figures. It contains: Figure S1: Microfluidic Assay for cell migration in 3D ECM; Figure S2: Macrophages increase cancer cell invasion rate in 3D ECM; macrophages increase cancer cell migration total speed but decrease directedness on 2D substrates; Figure S3: MMP activity is crucial for cancer cell migration, and macrophages enhance cancer cell MMP expression; Figure S4: The abilities of macrophages to enhance cancer cell migration dynamics and the expression of MMPs are due primarily to the paracrine factors secreted by macrophages; Figure S5: Neutralizing antibody blocking is specific to factors released by macrophages; Figure S6: Blocking TNFalpha and TGFbeta1 secreted by macrophages significantly reduced macrophage-induced MMP expression and macrophage-enhanced cancer cell invasion rate; Figure S7: TNFalpha and TGFbeta1 secreted by macrophages mostly accounts for macrophage-enhanced cancer cell migration total speed and directedness; Figure S8: TGFbeta1 has a larger effect on cancer cell migration total speed than TNFalpha. TNFalpha and TGFbeta1 synergistically increase cancer cell migration directedness; Figure S9: Cancer cell migration total speed is correlated with cancer cell expression of MT1-MMP, while cancer cell migration directedness is correlated with cancer cell expression of MMP1; Figure S10: Inhibiting both TNFalpha and TGFbeta1 in a mice model of breast tumor effectively reduces lung metastasis; Figure S11: TNFalpha and TGFbeta1 co-treatment synergistically up-regulate cancer cell MMP1 protein production and secretion. TGFbeta1 is mainly responsible for the increase in MT1-MMP protein production; Figure S12: TNFalpha and TGFbeta1 co-treatment synergistically enhance NF-κB nuclear localization in MDA-MB-435S cells. Fig. S1 and S2 is related to Fig. 1; Fig. S3 and S4 is related to Fig. 2; Fig. S5, S6, S7 is related to Fig. 3; Fig. S8-11 is related to Fig. 4; Fig. S12 is related to Fig. 6</p>