Journal: bioRxiv

Article Title: Estradiol alters actin and protrusion dynamics in endometriotic epithelial cells

doi: 10.1101/2025.05.14.654086

Figure Lengend Snippet: Experimental setup. (a) 12Z cells were transfected with a LifeAct-GFP plasmid via nucleofection (an electroporation-based method) and imaged over time using volumetric lattice lightsheet microscopy, which employs long thin beams to illuminate the sample with subcellular resolution. (b) A maximum intensity projection showing that the boundary of the actin cytoskeleton (LifeAct-GFP) coincides with the plasma membrane. (c) An example output maximum intensity projection from a lattice lightsheet microscopy time-lapse of a cell transfected with LifeAct-GFP with frames from t = 0, 1 min, 5 min, 10 min, and 60 min. Scalebars = 20 µm.

Article Snippet: The 12Z cell line was purchased from Applied Biological Materials (Cat. #T0764).

Techniques: Transfection, Plasmid Preparation, Electroporation, Microscopy, Clinical Proteomics, Membrane

Journal: bioRxiv

Article Title: Estradiol alters actin and protrusion dynamics in endometriotic epithelial cells

doi: 10.1101/2025.05.14.654086

Figure Lengend Snippet: 3D isosurfaces of 12Z cells obtained from lattice lightsheet imaging, with some notable cellular features (a-d) and representative time series (e-g) . ( a) Some cells exhibited actin waves at the leading edge of the cell and hair-like protrusions at the lagging edge of the cell (indicated by white arrows). Several cells also exhibited membrane ruffling (b) , 3D filopodial extensions (c, d) , and active protrusions (d) . The dynamics of the membrane ruffling and lamellipodia growth were captured (e) . Protrusion retraction (f, g) and formation (f) were also visualized. All scalebars = 20 µm.

Article Snippet: The 12Z cell line was purchased from Applied Biological Materials (Cat. #T0764).

Techniques: Imaging, Membrane

Journal: bioRxiv

Article Title: Estradiol alters actin and protrusion dynamics in endometriotic epithelial cells

doi: 10.1101/2025.05.14.654086

Figure Lengend Snippet: Effect of E2 on 12Z shape and morphodynamics after 15 minutes or 24 hours of treatment incubation. Circularity (a) , rate of change in circularity (b) , solidity (c) , and rate of change in solidity (d) over the duration of each time-lapse were extracted from binarized maximum intensity projections, and the means ± SEM are plotted. * P ≤ 0.05, ** P≤ 0.01.

Article Snippet: The 12Z cell line was purchased from Applied Biological Materials (Cat. #T0764).

Techniques: Incubation

Journal: bioRxiv

Article Title: Estradiol alters actin and protrusion dynamics in endometriotic epithelial cells

doi: 10.1101/2025.05.14.654086

Figure Lengend Snippet: Effect of E2 on 12Z morphodynamics after 15 min (top) or 24 hr (bottom) of treatment incubation. Circularity and solidity were tracked over the duration of each experiment and plotted (a, c) , along with the average change in each parameter normalized to initial value over time intervals of varying lengths (b, d) . The base time step Δt = 10 seconds.

Article Snippet: The 12Z cell line was purchased from Applied Biological Materials (Cat. #T0764).

Techniques: Incubation

Journal: bioRxiv

Article Title: Estradiol alters actin and protrusion dynamics in endometriotic epithelial cells

doi: 10.1101/2025.05.14.654086

Figure Lengend Snippet: Impact of E2 on actin optical flow alignment in 12Z cells. (a) Optical flow (OF) was calculated from actin fluorescence for all timepoints to determine how the actin is moving within the video. The optical flow vectors are displayed on top of one of the corresponding actin fluorescence images. (b) The actin optical flow alignment (OF alignment) is displayed for the cell in (a) . Higher values of optical flow alignment mean that the optical flow in that region is pointing in the same direction (see Materials and methods). Using the binarized masks from the average optical flow alignment was found inside the protrusions and the cell body. The mean protrusion optical flow alignment (c) and mean ratio of protrusion optical flow alignment to cell body optical flow alignment (d) are plotted ± SEM. * P ≤ 0.05, ** P≤ 0.01.

Article Snippet: The 12Z cell line was purchased from Applied Biological Materials (Cat. #T0764).

Techniques: Fluorescence

Journal: Cells

Article Title: Endometriosis and Cytoskeletal Remodeling: The Functional Role of Actin-Binding Proteins

doi: 10.3390/cells14050360

Figure Lengend Snippet: Summary of the involvement of ABP proteins in endometriosis. WB—Western blot, IHC—immunohistochemistry, PCR—polymerase chain reaction.

Article Snippet: , Eutopic endometrium of endometriosis-free controls in proliferative phase ( n = 10) , Ectopic endometrium from different localizations ( n = 23) 12Z cell line (endometriotic epithelial cells) , Sphingosine-1-phosphate receptor 3 expression is upregulated in endometriosis lesions, which correlates with EMT and fibrosis markers. Its effects are abolished by the EZR inhibitor NSC668394. , EZR and sphingosine-1-phosphate receptor 3 are functionally interconnected through their roles in cell signaling, cytoskeletal organization, and cellular migration, particularly in processes like cancer metastasis, inflammation, and tissue remodeling. Studies were partially conducted on 12Z cell line and are yet to be evaluated in primary cultures. , [ ] .

Techniques: Control, Expressing, Staining, Immunohistochemistry, Isolation, Phospho-proteomics, Knockdown, Western Blot, Migration, Activity Assay, Labeling, Cell Culture, Immunofluorescence, In Vitro, Diagnostic Assay, Activation Assay, Biomarker Discovery, Over Expression, In Vivo, Virus, Functional Assay, Gene Expression, Marker, Inhibition, Derivative Assay, Enzyme-linked Immunosorbent Assay, Fluorescence

Journal: Scientific Reports

Article Title: Apolipoprotein-B mRNA-editing complex 3B could be a new potential therapeutic target in endometriosis

doi: 10.1038/s41598-024-76589-2

Figure Lengend Snippet: HIF-1α, APOBEC3B, KRAS, and PIK3CA expressions were measured with RT-PCR in the 12Z endometriotic cell line at different concentrations of O 2 (20%O 2 , 5% O 2 , 1% O 2 ) for 48 h.

Article Snippet: 12Z immortalized human endometriotic cell line was obtained from Applied Biological Materials (Vancouver, Canada).

Techniques: Reverse Transcription Polymerase Chain Reaction

Journal: Scientific Reports

Article Title: Apolipoprotein-B mRNA-editing complex 3B could be a new potential therapeutic target in endometriosis

doi: 10.1038/s41598-024-76589-2

Figure Lengend Snippet: After transient transfection of the mock, control siRNA (siCon) and APOBEC3BsiRNA (siAPOBEC3B) into 12Z endometriotic cell line for 48 h: ( A ) RT-PCR analysis of the APOBEC3B expression levels in mock, siCon and siAPOBEC3B cells. ( B ) MTS assays of mock, siCon, and siAPOBEC3B cells. ( C ) Representative flow cytometric data for apoptosis levels in mock, siCon, and siAPOBEC3B cells. ( D ) RT-PCR analysis of the HIF-1α expression levels in mock, siCon, and siAPOBEC3B cells.

Article Snippet: 12Z immortalized human endometriotic cell line was obtained from Applied Biological Materials (Vancouver, Canada).

Techniques: Transfection, Control, Reverse Transcription Polymerase Chain Reaction, Expressing

Journal: Scientific Reports

Article Title: Apolipoprotein-B mRNA-editing complex 3B could be a new potential therapeutic target in endometriosis

doi: 10.1038/s41598-024-76589-2

Figure Lengend Snippet: After transient transfection of the mock, control siRNA (siCon) and APOBEC3BsiRNA (siAPOBEC3B) into 12Z endometriotic cell line for 48 h: ( A ) RT-PCR analysis of KRAS, PIK3CA, Caspase 7 and Caspase 8 expressions compared between mock, siCon and siAPOBEC3B cells. ( B ) Cell invasion assay compared between mock, siCon, and siAPOBEC3B transfected cells. ( C ) Cell migration assay compared between mock, siCon, and siAPOBEC3B transfected cells. Cell invasion and migration assays were performed in triplicate. ( D ) Cell growth in monolayers was compared between mock, siCon, and siAPOBEC3B transfected cells using DMEM/ham’s F12 medium supplemented with 10% FBS for 2, 4, and 6 days. The numbers represent data from triplicate experiments.

Article Snippet: 12Z immortalized human endometriotic cell line was obtained from Applied Biological Materials (Vancouver, Canada).

Techniques: Transfection, Control, Reverse Transcription Polymerase Chain Reaction, Invasion Assay, Cell Migration Assay, Migration