Fig. 4

Exosomal lncMMPA enhances macrophage-induced tumor progression. A Electron microscope observation of the morphology of TAMs-derived exosomes. Scale bar, 100 nm. B Western blot analysis of antigens (CD9, Alix and TSG101) in TAMs-derived exosomes (TAMs-exo). C The expression level of lncMMPA in the equal numbers of exosomes derived from MDMs (left) and TAMs (right) after treatment with RNase A alone or together with Triton X-100 was normalized to that of cel-miR-39-3p and shown as a ratio relative to the expression level in untreated exosomes derived from MDMs or TAMs. D Fluorescent staining was conducted in Hep3B cells following incubation with Dil-labeled (red) exosomes obtained from TAMs. Scale bar, 20 μm. E-H Co-culture of Hep3B cells with MDMs and pri-TAMs was done in Transwell systems for 6 d. Separately, Hep3B cells were treated with exosome from TAMs for 48 h. After these treatments, Hep3B cells were collected for the specified experiments. E Glucose consumption. F Lactate production. G ECAR. H Expression of GLUT1, HK2 and LDHA. I-L Hep3B cells were treated with exosomes from MDMs with or without overexpressing lncMMPA and then harvested for the specified experiments. I Glucose consumption. J Lactate production. K ECAR. L Expression of GLUT1, HK2 and LDHA. M, N Hep3B cells were injected orthotopically into the right flank of BALB/c nude mice. The mice were given intratumoral injection of the indicated exosomes from MDMs every 3 d. M Representative images of tumors in the xenografts. N Tumor growth curves. O-R Hep3B cells were treated with exosomes from TAMs with or without shlncMMPA transfection and then collected for the indicated experiments. O Glucose consumption. P Lactate production. Q ECAR. R Expression of GLUT1, HK2 and LDHA