Journal: bioRxiv

Article Title: Hyperactivated YAP1 Drives an Invasive EMT Subtype of Cervical Squamous Cell Carcinoma

doi: 10.64898/2026.01.18.700207

Figure Lengend Snippet: A ) Representative images showing the ureter obstruction in of Krt14 -rtTA;Tet-on-YAP S127A mice induced by doxycycline (DOX-treated, lower panel). Krt14 -rtTA;Tet-on-YAP S127A mice administered with water (H 2 O, upper panel) were used as control. Note the enlarged bladders caused by the ureteral obstruction due to tumorigenesis in DOX-treated Krt14 -rtTA;Tet-on-YAP S127A mice induced (n = 5-8/group). B ) Representative images showing the histology (H&E staining) of cervical epithelium in the Krt14 -rtTA;Tet-on-YAP S127A mice induced with doxycycline (DOX, lower panel) or water (H2O, upper panel). Arrowheads point to the invading site where transformed epithelial cells break the basal membrane and invade into the stromal layer of cervical tissues. Scale bar: 100 µm. C ) representative images showing the expression of YAP1 protein (in brown, detected by IHC) in cervical tissues of Krt14 -rtTA;Tet-on-YAP S127A mice induced with water as control. Scale bar: 200 µm. D ) representative images showing the expression of YAP1 protein (in brown, detected by IHC) in cervical tissues of Krt14 -rtTA;Tet-on-YAP S127A mice administered with doxycycline. Scale bar: or DOX (lower panel). A dash-lined red rectangle box was used to highlight the transformation zone (TZ) of mouse uterine cervix. Two blue-line squares in the transformation zone highlight the newly formed cancer tissues in the cervix of Dox-induced Krt14 -rtTA;Tet-on-YAP S127A mice. UT: uterine tubes; CVX: cervix. Scale bar: 200 µm. E ) Representative images showing expression of E-cadherin (in red, detected by fluorescent immunohistochemistry) in the cervical tissue of Krt14 -rtTA;Tet-on-YAP S127A mice administered with water (H2O, left) or doxycycline (Dox-treated, right). Carcinoma cells invaded into the stromal area, which express relatively low E-cadherin (CDH1 + ), were circled with a white dished-line. Scale bar: 100 µm.

Article Snippet: Krt14-rtTA mice (FVB-Tg(KRT14-rtTA) F42Efu/J, #008099, The Jackson Laboratory) were crossed with Tet-on-YAP S127A mice (a gift from Dr. Fernando Camargo’s lab, Boston Children’s Hospital) to generate Krt14-rtTA; Tet-on-YAP S127A mice, which express a constitutively active form of YAP1 (YAP S127A ) under the control of a Krt14-driven tetracycline regulatory element (TRE) ( ).

Techniques: Control, Staining, Transformation Assay, Membrane, Expressing, Immunohistochemistry

Journal: bioRxiv

Article Title: Hyperactivated YAP1 Drives an Invasive EMT Subtype of Cervical Squamous Cell Carcinoma

doi: 10.64898/2026.01.18.700207

Figure Lengend Snippet: A ) UMAP plot showing overview of 9 major cell types identified in cervical tissues in Krt14 -rtTA;Tet-on-YAP S127A mice administered with water (H2O) as control or doxycycline (DOX) for tumorigenesis. B ) Violin plots showing expression of marker genes for the identified cell clusters, including markers for Basal/Parabasal epithelial cell (EC) 1, Basal/Parabasal EC 2, Keratinocyte, Cancer cell, two myeloid cells (Myeloid_1 & Myeloid_2), Lymphocyte, Endothelial cell, and Stroma cells. C ) UMAP plots showing the alteration of cellular components in cervical tissues of Krt14 -rtTA;Tet-on-YAP S127A mice administered with water (H2O) as control or doxycycline (DOX) for induction of tumorigenesis. Arrow points to the newly formed carcinoma cells, while the arrowhead points to the accumulated myeloid 2 cells (tumor-associated macrophages) during early-stage of tumorigenesis. D ) Bar plots overlaid with a smoothed area curve showing dynamic alterations of major cellular components of cervical tissues of Krt14 -rtTA;Tet-on-YAP S127A mice during Doxycycline (DOX)-induced tumorigenesis. Krt14 -rtTA;Tet-on-YAP S127A mice treated with water (H 2 O) were used as control. *: significantly different from the control (H 2 O). Statistical differences were calculated with scCODA with FDR = 0.05.

Article Snippet: Krt14-rtTA mice (FVB-Tg(KRT14-rtTA) F42Efu/J, #008099, The Jackson Laboratory) were crossed with Tet-on-YAP S127A mice (a gift from Dr. Fernando Camargo’s lab, Boston Children’s Hospital) to generate Krt14-rtTA; Tet-on-YAP S127A mice, which express a constitutively active form of YAP1 (YAP S127A ) under the control of a Krt14-driven tetracycline regulatory element (TRE) ( ).

Techniques: Control, Expressing, Marker

Journal: bioRxiv

Article Title: Hyperactivated YAP1 Drives an Invasive EMT Subtype of Cervical Squamous Cell Carcinoma

doi: 10.64898/2026.01.18.700207

Figure Lengend Snippet: A & B ) Spatial transcriptomic plot showing that unsupervised clustering based on spatial transcriptomics data successfully captured the morphology of cervical tissue in Krt14 -rtTA;Tet-on-YAP S127A mice administered with water (H 2 O, panel A) and Doxycycline (DoX, panel B). The identified cell clusters are presented in different colors. C & D ) Spatial distribution of sc-RNAseq-identified cells in the cervical tissues of KY mice treated with water (H 2 O, panel C, as negative control) or doxycycline (DOX, panel D). The cells were identified using marker/reference genes derived from the SC-RNAseq analyses of the matched samples. The color codes used to annotate cells in panel C) and panel D) are same. The dash-border square highlights the area with the newly formed cancer cells. A zoom-in view was presented on the right to visualize cancer cells (red) and tumor-associated myeloid cells (bright green, myeloid_2) in the tumor region. E ) Spatial transcriptomic plots mapping the location of each individual cell type in cervical tissues in Krt14 -rtTA;Tet-on-YAP S127A mice treated with water as control (H 2 O, left) or doxycycline (DOX, right) to induce tumorigenesis. Cells were annotated using cell markers/references identified using sc-RNAseq analyses of the matched samples. Color intensity encodes the prediction score, corresponding to the confidence level of the cell-type assignment. F ) Spatial transcriptomic plots mapping the expression and location of YAP1 , Myc , Bcar1 and Cdkn2a genes in cervical tissues of Krt14 -rtTA;Tet-on-YAP S127A mice treated with water as control (H 2 O, left) or doxycycline (DOX, right) to induce tumorigenesis. The dash-border circle highlights the cancer region. Sequential color scales represent relative expression levels of dedicated genes.

Article Snippet: Krt14-rtTA mice (FVB-Tg(KRT14-rtTA) F42Efu/J, #008099, The Jackson Laboratory) were crossed with Tet-on-YAP S127A mice (a gift from Dr. Fernando Camargo’s lab, Boston Children’s Hospital) to generate Krt14-rtTA; Tet-on-YAP S127A mice, which express a constitutively active form of YAP1 (YAP S127A ) under the control of a Krt14-driven tetracycline regulatory element (TRE) ( ).

Techniques: Negative Control, Marker, Derivative Assay, Control, Expressing

Journal: bioRxiv

Article Title: Hyperactivated YAP1 Drives an Invasive EMT Subtype of Cervical Squamous Cell Carcinoma

doi: 10.64898/2026.01.18.700207

Figure Lengend Snippet: A ) UMAP plot derived from scRNAseq data showing density of YAP1 expression in cervical cells of KY mice administered with water as control (H 2 O) or Dox to induce cervical cancer. The dash-border circle highlights the cluster of cancer cells, which have the highest YAP1 transgene. B ) GSEA plot derived from scRNAseq data showing enrichment of YAP1 signature in cancer cells in Krt14 -rtTA;Tet-on-YAP S127A mice induced with doxycycline (DOX) compared with cervical epithelial cells in control mice (KY mice treated with H 2 O). C ) UMAP plot derived from scRNAseq data showing epithelial cell subtypes in cervical tissues in KY mice treated with water (H 2 O) as control or Dox to induce CVC. The dash-border circle highlights the cluster of cancer cells. B/P_EpC; basal/parabasal epithelial cells; K_EpCs: Keratinized cervical epithelial cells. D ) The pathways enriched in invasive cancer cells in Dox-induced KY mice when compared to cervical epithelial cells in control mice (KY mice treated with H2O as negative control). Color scales show adjusted P values. E ) Spatial transcriptomics plot showing density of EMT signature subtypes in cervical tissues in control (H 2 O) and Dox-induced KY mice. The dash-border circle highlights the cluster of cancer cells. Sequential color scale represents the signal density of EMT signature genes. The dash-border circle highlights the cancer region. F ) A bubble chart showing the upregulation of EMT signature genes in invasive cancer cells. The relative gene expression data of EMT signature genes in invasive cancer cells and control cervical epithelial cells were derived from single-cell RNA sequencing analyses of single cells isolated from cervical tissues from doxycycline-induced KY mice and non-induced KY mice (H 2 O+, used as negative control). The color represented average log2 fold changes (avg_log2FC) in gene expression levels of invasive cancer cells relative to non-induced control cervical epithelial cells (control EpCs-H2O+). The size marks the p values generated by non-parametric Wilcoxon rank sum test implemented in R package Seurat. The analysis was restricted to human EMT signature ortholog genes . Notably, 95% of EMT signature genes are upregulated in invasive cancer cells compared to non-induced control cervical epithelial cells.

Article Snippet: Krt14-rtTA mice (FVB-Tg(KRT14-rtTA) F42Efu/J, #008099, The Jackson Laboratory) were crossed with Tet-on-YAP S127A mice (a gift from Dr. Fernando Camargo’s lab, Boston Children’s Hospital) to generate Krt14-rtTA; Tet-on-YAP S127A mice, which express a constitutively active form of YAP1 (YAP S127A ) under the control of a Krt14-driven tetracycline regulatory element (TRE) ( ).

Techniques: Derivative Assay, Expressing, Control, Negative Control, Gene Expression, RNA Sequencing, Isolation, Generated

Journal: bioRxiv

Article Title: Hyperactivated YAP1 Drives an Invasive EMT Subtype of Cervical Squamous Cell Carcinoma

doi: 10.64898/2026.01.18.700207

Figure Lengend Snippet: A ) UMAP plot showing subtypes of myeloid cells in cervical tissue in Krt14 -rtTA;Tet-on-YAP S127A mice administered with water as control (H 2 O) or doxycycline (Dox) to induce cervical cancer. MDSC: Myeloid-derived suppressor cells; Mono/M: other myeloid cells; TRM: Tissue resident macrophages. B ) Violin plots showing expression of representative marker genes of the identified the major myeloid subtypes in A). C ) Dynamic alterations of myeloid cell subtypes in cervical tissues in control (H 2 O) and doxycycline-induced (DOX) Krt14-rtTA;Tet-on-YAPS 127A mice. *: significantly different when compared to control group. Statistical difference was performed with scCODA with FDR= 0.05. D ) Spatial transcriptomic plot showing the location of MDSCs (identified using matched scRNAseq samples as reference) subtypes in cervical tissues in control (H 2 O) and doxycycline-induced (DOX) Krt14 -rtTA;Tet-on-YAPS 127A mice. Sequential color scales represent the prediction score / confidence of cell type assignments. The dash-border circle highlights the cancer region. E ) The pathways enriched in MDSCs in the cervical tissues of Krt14 -rtTA;Tet-on-YAPS 127A mice induced with doxycycline (DOX) when compared with that of myeloid cells in cervical tissues of the control mice ( Krt14 -rtTA;Tet-on-YAPS 127A mice administered with H 2 O). Color scales show adjusted P values. F) The active ligand-receptor interactions between cancer cells and MDSCs. Magnitude is calculated as the arithmetic average of the mean expression of the ligand in the senders and the receptor in the receivers, using the minimum subunit expression for any multi-subunit complexes involved. P values were derived by random permutation of cell cluster labels implemented in Python package CellphoneDB. A p <1E-3 is considered specific. Please note that the primary interactions are associated with chemotaxis, immunosuppression, EMT and tumor invasiveness. G) Spatial distance and signal strength weighted density of ligand-receptor interactions between cancer cells and MDSCs in cervical tissues of control (H 2 O) and doxycycline-induced (DOX) Krt14 -rtTA;Tet-on-YAPS 127A mice. Please note that the primary interactions are associated with immunosuppressive, EMT and tumor invasive. Sequential color scales represent the signal density of a specific interaction between cancer cells and MDSCs in the indicated tissues.

Article Snippet: Krt14-rtTA mice (FVB-Tg(KRT14-rtTA) F42Efu/J, #008099, The Jackson Laboratory) were crossed with Tet-on-YAP S127A mice (a gift from Dr. Fernando Camargo’s lab, Boston Children’s Hospital) to generate Krt14-rtTA; Tet-on-YAP S127A mice, which express a constitutively active form of YAP1 (YAP S127A ) under the control of a Krt14-driven tetracycline regulatory element (TRE) ( ).

Techniques: Control, Derivative Assay, Expressing, Marker, Chemotaxis Assay