Fig. 3

The attenuating effect of SERPINB2 knockdown on the acetylsalicylic acid-induced suppression of the Akt or ERK1/2 signaling cascade. A schematic representation of the role of SERPINB2 as an upstream regulator of the acetylsalicylic acid-induced PI3K/Akt and FAK/ERK1/2 signaling cascades is shown (A). Endometrial stem cells were treated with or without acetylsalicylic acid (2.5 mM) for 15 min, and the subsequent changes in the phosphorylation (activation) levels of signaling molecules (i.e., Akt, PI3K, FAK, and ERK1/2) were assessed by western blotting (B-C). Endometrial stem cells were treated with acetylsalicylic acid (2.5 mM) alone or concomitantly transfected with a specific shRNA targeting SERPINB2; the subsequent changes in the phosphorylation states of these signaling molecules were evaluated by western blotting (D-E). The activation states (either activated or inactivated) of various Akt1 (GSE100752) (F) or MAPK1/3 (ERK1/3) (GSE129144/GSE76381) (G)-associated genes/ transcription factors in proliferating and nonproliferating cells were analyzed using the ingenuity pathway analysis (IPA) software. The GEO metadata were also analyzed to further investigate the contributions of the Akt and MAPK1/3 (ERK1/3) signaling pathways to various acetylsalicylic acid treatment conditions (H). β-actin was used as an internal control to normalize protein expression. All experiments were performed in triplicates. Significant differences are presented. *p < 0.05, **p < 0.005, and ***p < 0.001 (One-way ANOVA)