Fig. 5.

RFP14 specifically inhibits RAB25-dependent migration in cancer cells. a, b Representative time-dependent scratch-wound cell migration assay images a and quantification b of mock vector and RAB25-expressing HEY, MCF7, and MDA-MB231 cells. RAB25 expression results in significant enhancement (HEY, MCF7) or inhibition (MDA-MB231) of cell migration. These data are representative of triplicate biological replicates. c, d RFP 14 (10 μM) significantly impaired cell migration of RAB25-expressing d, but not vector control c, HEY cells. e RFP14 (10 μM), but not RFP32, significantly impaired cell migration of RAB25-expressing MCF7 cells. f Treatment of RAB25-expressing MDA-MB231 cells with RFP14, conversely, significantly increased cell migration relative to vehicle while the inactive control RFP32 remained unchanged compared to vehicle control. Representative scratch wound images are shown for RFP14, RFP32, and vehicle (DMSO) treatment conditions with overlaid arrows highlighting distance between cell fronts. Images and data shown represent mean ± s.e.m. of triplicate biological replicates. *P < 0.05; **P < 0.01, Student’s t-test. NS, not significant. Inset scale bars for HEY cells in a = 250 μm; MCF7 and MDA-MB231 cells in a = 100 μm. Inset scale bars for c, d, e, f = 250 μm