Fig. 1

Effect of PGE2 and pAAV/pAAV-GlyR1/3 on ERK phosphorylation and ATF-3 activation in F11 cells. A Time schedule of PGE2 (100 µM) administration and the time when cells were collected to measure ERK phosphorylation and ATF-3 activation. F11 cells were seeded into 6-well plates, and 24 h later, PGE2 was applied for 5, 15, 30 or 60 min. Finally, these treated cells were harvested for protein extraction. B Western blot images and quantitative evaluation of C p-ERK and D ATF-3 activation in PGE2-treated F11 cells. E Time schedule of F11 cells transfected with 2 μg pAAV, pAAV-GlyRα1 or pAAV-GlyRα3 for 48 h. Then, cell pellets were harvested for protein extraction. F Images of western blots and quantitative evaluation of G p-ERK and H ATF3 expression in pAAV-, pAAV-GlyR1 or pAAV-GlyRα3-transfected F11 cells are shown. I Time schedule to evaluate the effect of pAAV, pAAV-GlyRα1/3 transfection on PGE2 (100 µM)-induced ERK phosphorylation in F11 cells. F11 cells were transfected with 2 μg pAAV, pAAV-GlyRα1 or pAAV-GlyRα3 for 48 h. Serum-free medium replaced the initial medium and was incubated for another 24 h, and the cells were then treated with PGE2 for 60 min. The cells were harvested for J western blotting of phosphorylated ERK, and K quantification of phosphorylated ERK in F11 cells is shown. F11 cells without vector transfection or PGE2 treatment were used as controls. All the data are expressed as the fold change measured in from three to five independent experiments. *p < 0.05, **p < 0.01, and ***p < 0.001, one-way ANOVA