Fig. 4

AKT and ERK phosphorylation are implicated in the combined action of rofecoxib and exemestane. A Representative experiment out of three independent western blot analyses of pCREB, pERK and pAKT expression in Ist Mes1, Ist Mes2 and MPP89 cells treated with 35µM exemestane (EXE) or 35µM rofecoxib (ROF) or 35µM exemestane and 35µM rofecoxib combination (EXE + ROF) for 30 min. B Representative western blot analyses of COX-2 and CYP19A1 expression in Ist Mes1, Ist Mes2 and MPP89 cells treated with 35µM exemestane (EXE) or 35µM rofecoxib (ROF) or 35µM exemestane and 35µM rofecoxib combination (EXE + ROF) for 24 h. C The graphs represent the mean ± SD of three independent quantifications of protein band intensities normalized to the loading control and then in comparison to the untreated sample (relative band intensity). D The graph represents the mean ± SD of three independent cell survival rates after pre-incubation with MK-2206 and subsequently treated with exemestane (MK-EXE), or rofecoxib (MK + ROF) or exemestane and rofecoxib combination (MK-EXE + ROF) compared to untreated (100 % of cell alive). E Representative western blot analyses of pAKT and pERK expression in Ist Mes2 cells pre-incubated with MK-2206 and after treated with exemestane (MK-EXE), or rofecoxib (MK + ROF) or exemestane and rofecoxib combination (MK-EXE + ROF). F The graph represents the mean ± SD of three independent quantifications of protein band intensities normalized to the loading control. Statistically significant effects (paired Student t test P < 0.05) compared to CNTR *, EXE # or ROF § or MK-CNTR °