Fig. 3

FFA1 regulates the migratory and invasive capacity of ACHN pRCC cells. A, B The effects of FFA1 on the migration of ACHN pRCC cells were evaluated by wound scratch assay. All conditions are in the presence of MMC to inhibit proliferation and MMC had no effect alone (not shown). PMA (1 µM) was used as positive control while media without serum was used as a negative control (not shown). PMA treated wounds closed fully after 24 h (p < 0.05; d = 5.9, versus control), while AS2034178 (10 µM) treated cells had significant wider wounds than control cells (p < 0.01; d = 5.9, versus control). The effects of AS2034178 were fully inhibited by GW1100 (p < 0.01; d = 4.7, versus AS2034178 alone), while GW1100 alone also caused near-full closure of the wound. Statistical significance was determined by paired t-test. * denotes p < 0.05 and ** denotes p < 0.01 compared to the vehicle-treated control condition, while ## denotes p < 0.01 compared to the AS2034178-treated condition. C, D The role of FFA1 in migration was also confirmed using in vitro transwell cell migration assays. AS2034178 (10 µM) facilitated significantly lower migration compared to vehicle-treated controls (p < 0.01; d = 11.1, versus control), while GW1100 significantly inhibited this effect (p < 0.01; d = 6.7, versus AS2034178). GW1100 alone demonstrated significantly higher migration compared to control (p < 0.01; d = 2.7, versus control). Graphs depict combined replicate data from three independent experiments. Statistical significance was determined by paired t-test. ** denotes p < 0.01 versus the control condition, while ## denotes p < 0.01 versus the AS2034178-treated condition. E, F The invasive capacity of pRCC cells was evaluated by a Matrigel-coated transwell assay. AS2034178 (10 µM) facilitated significantly lower invasion through the matrix compared to vehicle-treated controls (p < 0.01; d = 17.3, versus control), and GW1100 significantly inhibited this effect (p < 0.01; d = 15.6, versus AS2034178). GW1100 alone demonstrated significantly higher migration compared to control (p < 0.01; d = 8.0, versus control). The graph depicts combined replicate data from three independent experiments. Statistical significance was determined by paired t-test. ** denotes p < 0.01 versus the control condition and ## denotes p < 0.01 versus the AS2034178-treated condition