Figure 3

Analysis of proteins in the supernatant of co-cultures of MSCs and activated TIL. The levels of selected proteins from culture supernatant of MSCs co-cultured with TIL and M38 melanoma, compared with MSCs co-cultured with control TIL and M28 melanoma. (A) Principal component analysis (PCA) shows that the expression pattern of MSCs co-cultured with TIL and M38 melanoma (green) is different from that of MSCs co-cultured with TIL and M28 melanoma (red) and MSC monocultures (blue). (B) Unsupervised hierarchical clustering analysis of the protein levels revealed that all of the MSCs co-cultured with TIL and M38 melanoma are in one cluster (green), and MSCs co-cultured with TIL and M28 melanoma (red) clustered together with MSC monoculture (blue). (C) The heat map of supernatant protein levels shows that M-CSF, IFNγ, IL-1α, IL-5, TNFα, GM-CSF have almost same-level expression in the co-culture supernatants of TIL+M38 and BMSC+TIL+M38, implying those factors were secreted by activated TIL, while CXCL1, IL-8, VCAM1, CCL2, IL-6, CXCL10, CCL5, CXCL9, CXCL11, CCL8, IL-12, and CXCL10 were secreted by MSCs. MSCs from one subject were tested at three time points and were tested twice.