Fig. 6
FFA1 regulates pRCC cell invasion through inhibition of EGFR and ERK. A FFA1 agonism inhibits phosphorylation of EGFR at Tyr1068 compared to vehicle-treated control. The graphical results shown are representative of three independent experiments quantifying phosphorylation of EGFR at Tyr1068. B FFA1 agonism inhibits phosphorylation of ERK1/2, respectively, compared to vehicle-treated control. The graphical results shown are representative of three independent experiments quantifying phosphorylation of ERK1/2. C, D FFA1 agonism inhibits EGF-induced EGFR phosphorylation and ERK1/2 phosphorylation. The results shown are representative of two independent experiments. Statistical significance was determined by paired t-test. * denotes p < 0.05 and ** denotes p < 0.01 versus the control condition. E, F EGF (10 ng/mL) stimulated significantly higher invasion through the extracellular Matrigel matrix compared to vehicle-treated controls (p < 0.001; d = 8.5, versus control). Meanwhile, cells treated with AG1478 (500 nM) alone demonstrated significantly lower invasion compared to control cells (p < 0.05; d = 1.5, versus control), and again, AG1478 inhibited the positive regulation of invasion induced by EGF to decrease invasion (p < 0.001; d = 8.7, versus EGF). AS2034178 (10 μM) significantly reduced the invasion of pRCC cells through the matrix (p < 0.0001; d = 17.7, versus control) and AS2034178 also partially inhibited the positive regulation of EGF-induced invasion (p < 0.05; d = 3.2, versus EGF). Statistical significance was determined by paired t-test. Graphs depict combined replicate data from four independent experiments each performed in triplicate. * denotes p < 0.05, *** denotes p < 0.001, and **** denotes p < 0.0001 versus the control condition, while ### denotes p < 0.001 versus the EGF-treated condition and # denotes p < 0.05 versus the AS2034178-treated condition