Figure 10.

Tumor cell migration, elevated by inflammatory CAF-derived factors, is mediated through cooperativity between Ras-activating receptors and G protein-coupled receptors (GPCR) that signal via Gαi. Human MSCs were exposed to persistent TNFα + IL-1β stimulation (concentrations as in Figure 1) or to vehicles, generally for 14–18 days. Cytokine-devoid CM were collected as described in Figure 7 and were administered to MCF-7 cells in the presence of (A) FTS, an inhibitor of Ras; (B) PTx, an inhibitor of Gαi; or (C) both inhibitors together. Control cells were treated by the vehicles of inhibitors. Inhibitor concentrations were selected based on the considerations described in the Materials and Methods section. Cell numbers and viability were not affected by the inhibitors. Then, kinetics analyses of wound closure assays were performed in IncuCyte^®, as described in Figure 9B. The results are presented as mean ± SEM of 8–9 replicates for each treatment. *** p < 0.001, for differences between tumor cells treated by CM of inflammation-derived CAFs and control CAFs. ### p < 0.001, for differences between tumor cells treated by the inhibitors and tumor cells that were not treated by the inhibitors. The results of a representative experiment out of n = 3 are presented.