Fig. 2

FFA1 regulates ACHN cell proliferation. A The FFA1/4 agonist GW9508 (10 µM) induced significant increases in cell proliferation on days 5 (p < 0.05; d = 2.4, versus control) and 6 (p < 0.0001; d = 3.3, versus control). This effect of GW9508 was significantly inhibited by the selective FFA1 antagonist GW1100 (10 µM) on days 5 (p < 0.0001; d = 6.3, versus GW9508) and 6 (p < 0.0001; d = 6.38, versus GW9508). Compared to the vehicle-treated control condition, GW1100 alone significantly inhibited cell proliferation at days 5 (p < 0.001; d = 9.9, versus control) and 6 (p < 0.0001; d = 4.9, versus control). B The selective FFA1 agonist AS2034178 significantly increased cell proliferation on days 5 (p < 0.05; d = 1.4, versus control) and 6 (p < 0.0001; d = 1.6, versus control) compared to vehicle-treated cells, and the overall effect of AS2034178 was noticeably greater than that of the dual FFA1/4 agonist GW9508. GW1100 completely inhibited the effects of AS2034178 at day 5 (p < 0.0001; d = 4.1, versus AS2034178) and 6 (p < 0.0001; d = 3.1, versus AS2034178) of growth, while again, when used alone, GW1100 significantly inhibited cell proliferation at days 5 (p < 0.001; d = 38.9, versus control) and 6 (p < 0.0001; d = 16.0, versus control). Differences between groups were evaluated by two-way analysis of variance (ANOVA) followed by Tukey’s multiple comparisons post-hoc test. Graphs depict combined replicate data from three independent experiments each performed in triplicate. * denotes p < 0.05, *** denotes p < 0.001, and **** denotes p < 0.0001 versus the control condition. #### denotes p < 0.0001 versus the AS2034178-treated condition