Fig. 4
VSSP-activated BMDMs induce senescence in prostate cancer cells in vitro. PCa cell lines were cultured with RPMI 1640 containing 10% heat-inactivated FBS (normal medium), conditioned medium from BMDMs (1 part of normal medium and 1 part of unstimulated BMDMs conditioned medium) or conditioned medium from VSSP-activated BMDMs (1 part of normal medium and 1 part of VSSP-activated BMDMs CM). Then the cells were analyzed for Crystal Violet assay and/or for β-Galactosidase activity assay (after 72 h of culture, fold change compared with normal medium). a Proliferation assay on TrampC1-shPTEN cells, Tramp-C1 and Ptenpc−/−;Trp53pc−/− primary cells. Quantification was performed as fold change respect of the control. b Quantification of β-Galactosidase positive TrampC1-shPTEN, Tramp-C1 and Ptenpc−/−;Trp53pc−/− primary cells after treatment with the CM. Quantification was performed as fold change respect of the control. c Representative images of β-Galactosidase staining on TrampC1-shPTEN cells, Tramp-C1 and Ptenpc−/−;Trp53pc−/− primary cells. Positive stained cells are depicted by arrows. Symbols indicates significant differences by Student t test (**p < 0.01). BMDMs bone marrow-derived macrophages