Fig 2: Activation of PI3K/AKT/mTOR signaling pathway by mAb B1C4 was blocked by the dual PI3K/mTOR inhibitor (NVP-BEZ235). (A, B) The viability of HePG2 and Jurkat cells after treatment with NVP-BEZ235. The data are presented as the mean ±S.D. (n=3). *p<0.05, **p<0.01 vs 0 nM NVP-BEZ235. (C) Influence of NVP-BEZ235 on the protein expression by Jurkat cells in co-culture system detected by Western blot. Densitometric values were analyzed using the AlphaEaseFC 4.0 software program. ß-Actin was used as an internal control. Jurkat cells co-cultured with HePG2 cells for 48 h were used as controls and protein expression was normalized against the control. The data are presented as the means ± S.D. (n=3). *p<0.05, **p<0.01 vs Jurkat (PHA)+HepG2 (IFN-?); #p<0.05 vs (PHA)+HepG2 (IFN-?)+mAb (B1C4). (D) Influence of the dual PI3K/mTOR inhibitor (NVP-BEZ235) on killing ability of Jurkat cells in co-culture system detected by MTT assay. The data are presented as the means ± S.D. (n=3). **p<0.01 vs (PHA)+HepG2 (IFN-?); ##p<0.01 vs Jurkat (PHA)+HepG2 (IFN-?)+mAb (B1C4).

Fig 3: Alterations in the expression of PI3K and AKT in adenomyosis and FAK siRNA-treated cells. (A) RT-qPCR analysis of the mRNA expression levels of FAK, PI3K and AKT in cells of the adenomyosis and control groups. (B) Protein expression levels of FAK, PI3K and AKT in endometrial cells of the adenomyosis and control group were detected by western blotting. (C) Relative protein expression levels of FAK, PI3K and AKT in endometrial cells of the adenomyosis and control groups. (D) RT-qPCR analysis of the mRNA expression levels of FAK, PI3K and AKT in the FAK siRNA and negative control groups. (E and F) Western blot analysis of protein expression levels of FAK, PI3K and AKT in the FAK siRNA and negative control groups. The mRNA and protein expression levels were normalized to GAPDH; *P<0.05 and **P<0.01 vs. control group. AKT, protein kinase B; FAK, focal adhesion kinase; PI3K, phosphoinositide 3-kinase; RT-qPCR, reverse transcription-quantitative polymerase chain reaction; siRNA, small interfering RNA.

Fig 4: The expression of PTEN, p-Akt, Akt, p-mTOR, mTOR, Cleaved caspase-9, Cleaved caspase-3 and Bax in Jurkat cells determined by immunofluorescence staining. (A) Proteins stained by indirect fluorescence with secondary antibodies conjugated to Alexa Fluor-488 (green). Nuclei were counterstained with DAPI (blue). A representative image (n=3) is shown. Images were collected using a Leica TCS SP5 confocal imaging system. (B) The bar graphs show the fluorescence intensity of the proteins. The data are presented as the mean ± S.D. (n=10). *p<0.05, **p<0.01 vs Jurkat (PHA); ##p<0.01 vs Jurkat (PHA)+HepG2 (IFN-?).

Fig 5: The expressions of PTEN, p-Akt, Akt, p-mTOR, mTOR, Bax, Cleaved caspase-3 and Cleaved caspase-9 in Jurkat cells determined by Western blot. Densitometric values were analyzed using the AlphaEaseFC 4.0 software program. ß-Actin was used as an internal control. Jurkat cells activated by 2 µg/mL PHA for 48 h were used as controls and protein levels were normalized against the control. The data are presented as the means ± S.D. (n=3). *p<0.05, **p<0.01 vs Jurkat (PHA); #p< 0.05, ##p< 0.01 vs Jurkat (PHA)+HepG2 (IFN-?).