Fig. 3

Protein expression and functional detection of FFAR1/GPR40 in BUVEC monolayers. a Western blot of bovine FFAR1/GPR40. Total protein extract from LoVo cells was used as the positive control for two BUVEC isolates. 50 μg of total protein was loaded onto a 6 % polyacrylamide gel. b FFAR1/GPR40 immunofluorescence. Left image, nuclear signals of cells incubated with propidium iodide (PI); inset image: only the secondary antibody was used, as the negative control, bar = 20 μm. Middle image, FFAR1/GPR40 detected with rabbit anti-GPR40 antiserum and an anti-rabbit IgG secondary antibody. Right image, nuclear signals and GPR40 images are superimposed, bar = 20 nm. c Representative time course of the Fura-2/AM ratio in BUVECs exposed to 1 % DMSO or 100 μM TAK-875 at the time indicated by the arrow. d Cells were exposed to DMSO or TAK-875. The area under the curve (AUC) was calculated during the first 100 s after exposure to the stimulus. The data are expressed as the means ± SEM of at least three independent experiments. **p < 0.01, ***p < 0.005