Figure 4. Hyperactivation of YAP1 in Cervical Epithelial Cells Inactivates Signaling Pathways Involved in Type I Interferon Production In vitro and In vivo.
(A) Representative images from in vitro experiments showing expression and location of YAP1 and IRF3 in normal and “wound” area of cultured hCerECs. YAP1 and IRF3 proteins were visualized using an Alexa 488 (green)-conjugated secondary antibody. Nuclei were stained with DAPI (blue). Scale bar, 20 μm.
(B and C) Quantitative data showing mRNA levels of TLR2 and TLR4 (B), and their adapter proteins MYD88 and TRIF (C) in hCerEC-MX (control) and hCerEC-YAPS127A cells. Each bar represents mean ± SEM (n = 4). **p < 0.01 and ***p < 0.001.
(D and E) Quantitative data showing mRNA levels of IRF1 and IRF7 in hCerEC-MX (control) and hCerEC-YAPS127A cells (D) and in control (CTRL) and YAP1-knockdown hCerECs (siYAP) (E). Each bar represents the mean ± SEM (n = 4). **p < 0.01 and ***p < 0.001.
(F) Representative images showing the expression and location of YAP1 and IRF3 in hCerEC-MX (control), hCerEC-YAP, and hCerEC-YAPS127A cells. YAP1 and IRF3 proteins were visualized using an Alexa 488 (green)-conjugated secondary antibody. Nuclei were stained with DAPI (blue). Scale bar, 20 μm.
(G) RT-PCR analyses showing expressions of YAP1, TLR2, MYD88, and TBK1 mRNA in cervical tissues from KRT14-rtTA control (CTRL) and KRT14-YAPS127A mice. Each bar represents the mean ± SEM (n = 4). **p < 0.01 and ***p < 0.001.
(H) Representative images showing expression of YAP1, MYD88, and TBK1 proteins in cervical tissues from KRT14-rtTA control (CTRL) and KRT14-YAPS127A mice. Protein expression was determined using peroxidase-based immunohistochemistry. The nuclei were counterstained with hematoxylin. Note upregulation of YAP1 and downregulation of MYD88 and TBK1 proteins in the epithelium of KRT14-YAPS127A transgenic mice. Scale bar: 250 μm.