FIGURE 7.

JAM-C S-palmitoylation affects cell migration. A549 cells were transfected with FLAG-tagged JAM-C WT or the CCSS mutant for 24 h and then cultured in RPMI 1640 serum-free medium for 14 h. The cell migration assay was then performed in a 24-well Transwell plate with 8-μm polycarbonate sterile membranes. A total of 3.5 × 10⁴ cells in 200 μl of RPMI 1640 serum-free medium were plated into each upper chamber and placed in wells containing 600 μl of RPMI 1640 medium supplemented with 10% FBS. After 24 h, cells on the upper surface were detached with a cotton swab. The chambers were fixed, and cells in the lower filter were stained with 0.1% crystal violet for 15 min and counted. The quantified results were calculated by counting three random fields of migrated cells. The control cells were transfected with an empty pCMV4a vector. A, representative images of migrated cells from three independent experiments are shown. B, the number of migrated cells per field was quantified and normalized by the value of the control (n = 3; error bars represent S.D.). *, p < 0.05; **, p < 0.01; ***, p < 0.001.