Notch and PlexinD1 signaling regulate prostate cancer cell migration.
(A) Wound healing assay in PC3 cells treated with gamma secretase inhibitors—DAPT and RO4929097 (as in Fig 3C). Wound closure (24 hours from scratch) was quantified relative to wound width at start time. (B, C) The migration of PC3 cells stably expressing shScr, shNotch1, shPlexinD1 was analyzed in wound healing assays, as above. Panel B shows the quantification of wound sizes at the end of the experiment. (D) The migration of PC3 cells stably expressing shScr, shPlexinD1 and shNotch1 was assayed in overnight Boyden chamber experiments with transwell inserts. Bar graphs indicate mean values ± SD (normalized to controls). (E) PC3 cells overexpressing Dll1-Fc and Jag1-Fc were analyzed in Boyden chamber experiment, as above. (F) PC3 cells transduced to express an autocrine p61-Sema3E circuit were analyzed in Boyden chamber experiments, as above. (G) PC3 cells stably expressing shSema3E compared to shScr were analyzed in Boyden chamber experiments, as above. Mean ± SD is shown in all graphs; values were normalized to respective controls.