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. Author manuscript; available in PMC: 2016 Oct 4.

Published in final edited form as: Oncogene. 2016 Apr 4;35(38):5056–5069. doi: 10.1038/onc.2016.67

CD82 suppresses melanoma invasiveness and migration by modulating U2AF2-dependent CD44 isoform switching. (A) SK-Mel-25 cells that were transfected with vector or U2AF2 were cotransfected with shCD44s or shCD44v8-10. (B) SK-Mel-25 cells that were transfected with scrambled or CD82 siRNA were cotransfected with shU2AF2 or shCD44v8-10. (C, E) Lu1205M cells that were transfected with scrambled or U2AF2 shRNA were cotransfected with CD44s or CD44v8-10. (D, F) Lu1205M cells that were transfected with vector or CD82 were cotransfected with U2AF2 or CD44v8-10. (A-D) SK-Mel-25 or Lu1205M cells were tested for static migration abilities in transwell system. HUVEC monolayer was seeded on transwell insert membrane. The downward side of the membrane was stained with crystal violet and migrated cells were counted using an inverted microscope. Image magnification: 200X. Values are shown as mean±SEM from 12 fields from each of three independent experiments. *p<0.05. Bar=50 μm. (E-F) Lu1205M migration potentials under flow conditions were assessed with flow migration assays which were carried out for 4 hr at shear stresses of 0.625 and 2 dyn/cm². The migrated cells were stained and counted. Values are shown as mean±SEM from 12 fields from each of three independent experiments. *p<0.05.

CD82 suppresses melanoma invasiveness and migration by modulating U2AF2-dependent CD44 isoform switching. (A) SK-Mel-25 cells that were transfected with vector or U2AF2 were cotransfected with shCD44s or shCD44v8-10. (B) SK-Mel-25 cells that were transfected with scrambled or CD82 siRNA were cotransfected with shU2AF2 or shCD44v8-10. (C, E) Lu1205M cells that were transfected with scrambled or U2AF2 shRNA were cotransfected with CD44s or CD44v8-10. (D, F) Lu1205M cells that were transfected with vector or CD82 were cotransfected with U2AF2 or CD44v8-10. (A-D) SK-Mel-25 or Lu1205M cells were tested for static migration abilities in transwell system. HUVEC monolayer was seeded on transwell insert membrane. The downward side of the membrane was stained with crystal violet and migrated cells were counted using an inverted microscope. Image magnification: 200X. Values are shown as mean±SEM from 12 fields from each of three independent experiments. *p<0.05. Bar=50 μm. (E-F) Lu1205M migration potentials under flow conditions were assessed with flow migration assays which were carried out for 4 hr at shear stresses of 0.625 and 2 dyn/cm². The migrated cells were stained and counted. Values are shown as mean±SEM from 12 fields from each of three independent experiments. *p<0.05.