Fig. 2.

Effects of 27HC are mediated by its cancer-cell extrinsic effects on the host. a Mice were pretreated with placebo or 27HC (20 mg/kg) daily for 5 days, at which point Met1 cells expressing iRFP were grafted (i.v.) and treatment ceased. At day 28, lungs were harvested, imaged and iRFP fluorescence quantified. Experimental timeline outlined above. Representative images of iRFP-expressing Met1 colonies (depicted as black) within the lungs are presented above quantified data (placebo N = 5, 27HC N = 4, scale bar indicates 5 mm). b Mice were grafted (i.v.) with Met1 cells expressing iRFP on day 0. On day 5, mice were treated daily with placebo or 27HC for either 5 days or for the remainder of the study (chronic 27HC). At day 28, lungs were harvested, imaged and iRFP fluorescence quantified. Experimental timeline outlined above quantified data (placebo N = 7, 5 days 27HC N = 7, chronic 27HC N = 6). c Pretreatment of naive mice (regime outlined in a) with the CYP27A1 inhibitor GW297X (100 mg/kg) reduces colonization of lung by E0771 cells, which is reversed by co-pretreatment with 27HC (N = 5/group). d Pretreatment of mice with GW297X (as in c) results in decreased Met1 colonies. For this experiment, mice were pretreated with placebo or GW297X (100 mg/kg) for 5 days at which point Met1 cells expressing iRFP were grafted (i.v.) and treatment ceased. Five days post-graft, lungs were harvested, imaged, and fluorescence quantified (placebo N = 9, GW297X N = 10). Results are depicted as mean +/− SEM. Lines and asterisks denote statistical differences between groups (p < 0.05). a, d: Unpaired two-tailed student’s t-test. b, c: One-way ANOVA followed by a Student Newman-Keuls multiple comparison test