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human breast cancer mda mb 231 cell lines (ATCC)

Name: human breast cancer mda mb 231 cell lines
Brand: ATCC
Rating: 99 (27200 reviews)

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ATCC human breast cancer mda mb 231 cell lines
Human Breast Cancer Mda Mb 231 Cell Lines, supplied by ATCC, used in various techniques. Bioz Stars score: 99/100, based on 27200 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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Average 99 stars, based on 27200 article reviews

human breast cancer mda mb 231 cell lines - by Bioz Stars, 2026-02

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(A) Diagram of the breast cancer–on–a–chip platform used to simulate physiological laminar shear stress. (B) Time-course analysis of tumor-cell viability under laminar shear flow. MDA-MB-231 cells were uninfected or infected with wild-type, ΔdnaK, or ΔdnaK-C Staphylococcus xylosus. Cell viability was quantified at the indicated time points using a live/dead fluorescence assay and normalized to uninfected cells at 0 h. Data represent means ± SEM from three independent microfluidic experiments. (C) Quantification of tumor-cell viability at 4 h of shear-stress exposure. Each data point represents an independent microfluidic experiment. Statistical significance was determined by one-way ANOVA with Tukey’s post hoc test. p < 0.05; ns, not significant.

Journal: PLOS One

Article Title: DnaK supports intracellular persistence of Staphylococcus xylosus and confers mechanical resilience to a human breast cancer cell line

doi: 10.1371/journal.pone.0341069

Figure Lengend Snippet: (A) Diagram of the breast cancer–on–a–chip platform used to simulate physiological laminar shear stress. (B) Time-course analysis of tumor-cell viability under laminar shear flow. MDA-MB-231 cells were uninfected or infected with wild-type, ΔdnaK, or ΔdnaK-C Staphylococcus xylosus. Cell viability was quantified at the indicated time points using a live/dead fluorescence assay and normalized to uninfected cells at 0 h. Data represent means ± SEM from three independent microfluidic experiments. (C) Quantification of tumor-cell viability at 4 h of shear-stress exposure. Each data point represents an independent microfluidic experiment. Statistical significance was determined by one-way ANOVA with Tukey’s post hoc test. p < 0.05; ns, not significant.

Article Snippet: The human breast cancer cell line MDA-MB-231 was obtained from ATCC and cultured in DMEM supplemented with 10% fetal bovine serum (Gibco), 1% penicillin-streptomycin, and 2 mM L-glutamine.

Techniques: Shear, Infection, Fluorescence

Characterization of YAP/β-catenin expression and proliferation rate in breast cancer cells when subject to different stiffness (A) Random field of view images were taken under brightfield with a magnification of 20× of MCF-7 and MDA-MB-231 cells. Scale bars = 100 μm (B) Proliferation rates of MCF-7 and MDA-MB-231 cells on the substrate with different stiffness were measured using alamarBlue Cell viability reagent over the course of 8 days. Data represent mean ± SD. n = 3 wells per condition. Statistical significance was assessed using one-way ANOVA followed by the Tukey post hoc test. ∗ p < 0.05, ∗∗∗∗ p < 0.0001. (C) Western blot results of YAP and β-catenin in MCF-7 and MDA-MB-231 cells that were subject to 2 kPa and 32 kPa substrate. Protein expression of β-catenin, YAP, and pYAP was analyzed with GAPDH used for normalization.

Journal: iScience

Article Title: A stiffness-gated YAP-β-catenin axis orchestrates AXIN2 expression in metastatic breast cancer

doi: 10.1016/j.isci.2025.114405

Figure Lengend Snippet: Characterization of YAP/β-catenin expression and proliferation rate in breast cancer cells when subject to different stiffness (A) Random field of view images were taken under brightfield with a magnification of 20× of MCF-7 and MDA-MB-231 cells. Scale bars = 100 μm (B) Proliferation rates of MCF-7 and MDA-MB-231 cells on the substrate with different stiffness were measured using alamarBlue Cell viability reagent over the course of 8 days. Data represent mean ± SD. n = 3 wells per condition. Statistical significance was assessed using one-way ANOVA followed by the Tukey post hoc test. ∗ p < 0.05, ∗∗∗∗ p < 0.0001. (C) Western blot results of YAP and β-catenin in MCF-7 and MDA-MB-231 cells that were subject to 2 kPa and 32 kPa substrate. Protein expression of β-catenin, YAP, and pYAP was analyzed with GAPDH used for normalization.

Article Snippet: MDA-MB-231 human breast cancer cell line , ATCC , HTB-26; RRID: CVCL_0062.

Techniques: Expressing, Western Blot

YAP controls actin cytoskeletal organization and polymerization in a stiffness-dependent manner in metastatic breast cancer cells (A) MCF-7 and MDA-MB-231 cells were transfected with non-targeting control siRNAs (control) or siRNA against YAP (siYAP) for 48 h. Control group was treated with LatA at a concentration of 0.4 μM for 1 h (LatA). After the fixation, permeabilization, and blocking, the cells were stained for F-actin and G-actin. Images were taken using Cytation5 under 20× magnification. Scale bars = 50 μm (B) The quantification of the F/G actin ratio in MCF-7 cells (left) and MDA-MB-231 cells (right) from the images in A. Image analysis was performed using BioTek Gen5 Software, and bar graphs were created using GraphPad Prism. Data represent mean ± SD. n = individual cells pooled from 5 randomly acquired images per condition. Statistical significance was assessed using one-way ANOVA followed by the Tukey post hoc test. ns: not significant, ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001, ∗∗∗∗ p < 0.0001.

Journal: iScience

Article Title: A stiffness-gated YAP-β-catenin axis orchestrates AXIN2 expression in metastatic breast cancer

doi: 10.1016/j.isci.2025.114405

Figure Lengend Snippet: YAP controls actin cytoskeletal organization and polymerization in a stiffness-dependent manner in metastatic breast cancer cells (A) MCF-7 and MDA-MB-231 cells were transfected with non-targeting control siRNAs (control) or siRNA against YAP (siYAP) for 48 h. Control group was treated with LatA at a concentration of 0.4 μM for 1 h (LatA). After the fixation, permeabilization, and blocking, the cells were stained for F-actin and G-actin. Images were taken using Cytation5 under 20× magnification. Scale bars = 50 μm (B) The quantification of the F/G actin ratio in MCF-7 cells (left) and MDA-MB-231 cells (right) from the images in A. Image analysis was performed using BioTek Gen5 Software, and bar graphs were created using GraphPad Prism. Data represent mean ± SD. n = individual cells pooled from 5 randomly acquired images per condition. Statistical significance was assessed using one-way ANOVA followed by the Tukey post hoc test. ns: not significant, ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001, ∗∗∗∗ p < 0.0001.

Article Snippet: MDA-MB-231 human breast cancer cell line , ATCC , HTB-26; RRID: CVCL_0062.

Techniques: Transfection, Control, Concentration Assay, Blocking Assay, Staining, Software

Enhanced translocation of YAP and β-catenin to the nucleus occurs in response to a more rigid substrate in metastatic breast cancer cells (A) The intracellular distribution of β-catenin and YAP in MCF-7 and MDA-MB-231 cells, subjected to the substrates of 2 kPa and 32 kPa, was examined through immunofluorescent staining. Scale bars = 100 μm (B) YAP (top) and β-catenin (bottom) nuclear translocation was measured using BioTek Gen5 Software, and results were generated using GraphPad Prism. Data represent mean ± SD. n = individual cells pooled from 15 images (5 fields × 3 wells) per condition. Statistical significance was assessed using one-way ANOVA followed by the Tukey post hoc test. ns: not significant, ∗∗∗∗ p < 0.0001.

Journal: iScience

Article Title: A stiffness-gated YAP-β-catenin axis orchestrates AXIN2 expression in metastatic breast cancer

doi: 10.1016/j.isci.2025.114405

Figure Lengend Snippet: Enhanced translocation of YAP and β-catenin to the nucleus occurs in response to a more rigid substrate in metastatic breast cancer cells (A) The intracellular distribution of β-catenin and YAP in MCF-7 and MDA-MB-231 cells, subjected to the substrates of 2 kPa and 32 kPa, was examined through immunofluorescent staining. Scale bars = 100 μm (B) YAP (top) and β-catenin (bottom) nuclear translocation was measured using BioTek Gen5 Software, and results were generated using GraphPad Prism. Data represent mean ± SD. n = individual cells pooled from 15 images (5 fields × 3 wells) per condition. Statistical significance was assessed using one-way ANOVA followed by the Tukey post hoc test. ns: not significant, ∗∗∗∗ p < 0.0001.

Article Snippet: MDA-MB-231 human breast cancer cell line , ATCC , HTB-26; RRID: CVCL_0062.

Techniques: Translocation Assay, Staining, Software, Generated

YAP knockdown enhances β-catenin nuclear translocation on soft, but not stiff, substrates in metastatic breast cancer cells (A) The analysis of YAP and β-catenin expression in MDA-MB-231 cells upon YAP knockdown via immunofluorescent staining. Scale bars = 100 μm (B) The quantitative determination of nuclear translocation of β-catenin (top graph) and YAP (bottom graph) in MDA-MB-231 cells that were subjected 2 kPa and 32 kPa following the transfection of non-targeting siRNA (control) or siRNA against YAP (siYAP). n = 3 wells per condition. Data represent mean ± SD. Statistical significance was assessed using one-way ANOVA followed by the Tukey post hoc test. ns: not significant, ∗ p < 0.05.

Journal: iScience

Article Title: A stiffness-gated YAP-β-catenin axis orchestrates AXIN2 expression in metastatic breast cancer

doi: 10.1016/j.isci.2025.114405

Figure Lengend Snippet: YAP knockdown enhances β-catenin nuclear translocation on soft, but not stiff, substrates in metastatic breast cancer cells (A) The analysis of YAP and β-catenin expression in MDA-MB-231 cells upon YAP knockdown via immunofluorescent staining. Scale bars = 100 μm (B) The quantitative determination of nuclear translocation of β-catenin (top graph) and YAP (bottom graph) in MDA-MB-231 cells that were subjected 2 kPa and 32 kPa following the transfection of non-targeting siRNA (control) or siRNA against YAP (siYAP). n = 3 wells per condition. Data represent mean ± SD. Statistical significance was assessed using one-way ANOVA followed by the Tukey post hoc test. ns: not significant, ∗ p < 0.05.

Article Snippet: MDA-MB-231 human breast cancer cell line , ATCC , HTB-26; RRID: CVCL_0062.

Techniques: Knockdown, Translocation Assay, Expressing, Staining, Transfection, Control

β-catenin nuclear translocation following YAP knockdown is regulated by cell density in metastatic breast cancer cells (A) YAP and β-catenin in MDA-MB-231 cells at low, medium, and high confluency after YAP knockdown. Low confluency (30% confluency), medium confluency (50% confluency), and high confluency (80% confluency). MDA-MB-231 cells that were subject to 2 kPa and 32 kPa substrates were transfected with non-targeting siRNA (control) or siRNA against YAP (siYAP) before the immunofluorescent staining for YAP and β-catenin. Scale bars = 100 μm (B) The percentages of the cell population that show β-catenin nuclear translocation in A were measured in Agilent BioTek Gen 5 and plotted in GraphPad Prism. Data represent mean ± SD. n = 3 wells per condition. Statistical significance was assessed using one-way ANOVA followed by the Tukey post hoc test. ns: not significant, ∗ p < 0.05, ∗∗ p < 0.01. (C) The analysis of β-catenin and its active isoform expression in MCF-7 and MDA-MB-231 cells on 2 kPa and 32 kPa substrates following YAP knockdown. Total protein amount was used for normalization, and bar graphs were generated using GraphPad Prism. Data represent mean ± SD. n = 3 wells per condition. Statistical significance was assessed using one-way ANOVA followed by the Tukey post hoc test. ns: not significant.

Journal: iScience

Article Title: A stiffness-gated YAP-β-catenin axis orchestrates AXIN2 expression in metastatic breast cancer

doi: 10.1016/j.isci.2025.114405

Figure Lengend Snippet: β-catenin nuclear translocation following YAP knockdown is regulated by cell density in metastatic breast cancer cells (A) YAP and β-catenin in MDA-MB-231 cells at low, medium, and high confluency after YAP knockdown. Low confluency (30% confluency), medium confluency (50% confluency), and high confluency (80% confluency). MDA-MB-231 cells that were subject to 2 kPa and 32 kPa substrates were transfected with non-targeting siRNA (control) or siRNA against YAP (siYAP) before the immunofluorescent staining for YAP and β-catenin. Scale bars = 100 μm (B) The percentages of the cell population that show β-catenin nuclear translocation in A were measured in Agilent BioTek Gen 5 and plotted in GraphPad Prism. Data represent mean ± SD. n = 3 wells per condition. Statistical significance was assessed using one-way ANOVA followed by the Tukey post hoc test. ns: not significant, ∗ p < 0.05, ∗∗ p < 0.01. (C) The analysis of β-catenin and its active isoform expression in MCF-7 and MDA-MB-231 cells on 2 kPa and 32 kPa substrates following YAP knockdown. Total protein amount was used for normalization, and bar graphs were generated using GraphPad Prism. Data represent mean ± SD. n = 3 wells per condition. Statistical significance was assessed using one-way ANOVA followed by the Tukey post hoc test. ns: not significant.

Article Snippet: MDA-MB-231 human breast cancer cell line , ATCC , HTB-26; RRID: CVCL_0062.

Techniques: Translocation Assay, Knockdown, Transfection, Control, Staining, Expressing, Generated

β-catenin nuclear translocation upon YAP depletion supports cell proliferation and migration in metastatic breast cancer cells in a stiffness-dependent manner (A) Proliferation analysis of MCF-7 and MDA-MB-231 cells following YAP knockdown. Both MCF-7 cells and MDA-MB-231 cells were transfected with non-targeting siRNA (control), siRNA against YAP (siYAP), or siRNA against β-catenin (siβ-catenin). Then alamarBlue assay was performed for a period of 7 or 10 days at each time point for both MCF-7 cells and MDA-MB-231 cells. Then the data were generated using GraphPad Prism. Data represent mean ± SD. n = 3 wells per condition. (B) Cell migration analysis of MDA-MB-231 cells that were cultured on 2 kPa or 32 kPa substrates before the transfection with non-targeting siRNA (control), or siRNA against YAP (siYAP), or siRNA against YAP and siRNA against β-catenin (siYAP and siβ-catenin). Then the gap closure assay was performed, and the gap areas were imaged using Cytation5 under 4× magnification at 0 h and 20 h. Scale bars = 1000 μm (C) Quantitative analysis of the cell migration assay in B was conducted using ImageJ, and the results were generated using GraphPad Prism. Data represent mean ± SD. n = 3 wells per condition. Statistical significance was assessed using one-way ANOVA followed by the Tukey post hoc test. ns: not significant, ∗ p < 0.05.

Journal: iScience

Article Title: A stiffness-gated YAP-β-catenin axis orchestrates AXIN2 expression in metastatic breast cancer

doi: 10.1016/j.isci.2025.114405

Figure Lengend Snippet: β-catenin nuclear translocation upon YAP depletion supports cell proliferation and migration in metastatic breast cancer cells in a stiffness-dependent manner (A) Proliferation analysis of MCF-7 and MDA-MB-231 cells following YAP knockdown. Both MCF-7 cells and MDA-MB-231 cells were transfected with non-targeting siRNA (control), siRNA against YAP (siYAP), or siRNA against β-catenin (siβ-catenin). Then alamarBlue assay was performed for a period of 7 or 10 days at each time point for both MCF-7 cells and MDA-MB-231 cells. Then the data were generated using GraphPad Prism. Data represent mean ± SD. n = 3 wells per condition. (B) Cell migration analysis of MDA-MB-231 cells that were cultured on 2 kPa or 32 kPa substrates before the transfection with non-targeting siRNA (control), or siRNA against YAP (siYAP), or siRNA against YAP and siRNA against β-catenin (siYAP and siβ-catenin). Then the gap closure assay was performed, and the gap areas were imaged using Cytation5 under 4× magnification at 0 h and 20 h. Scale bars = 1000 μm (C) Quantitative analysis of the cell migration assay in B was conducted using ImageJ, and the results were generated using GraphPad Prism. Data represent mean ± SD. n = 3 wells per condition. Statistical significance was assessed using one-way ANOVA followed by the Tukey post hoc test. ns: not significant, ∗ p < 0.05.

Article Snippet: MDA-MB-231 human breast cancer cell line , ATCC , HTB-26; RRID: CVCL_0062.

Techniques: Translocation Assay, Migration, Knockdown, Transfection, Control, Alamar Blue Assay, Generated, Cell Culture, Cell Migration Assay

Myosin II activity facilitates YAP and β-catenin nuclear translocation, while intact actin filaments restrict β-catenin nuclear localization (A) MDA-MB-231 cells were transfected with non-targeting siRNA (control), siRNA against YAP (siYAP). Control or siYAP cells were treated with Blebbistatin at a concentration of 50 μM for 30 min or with LatA at a concentration of 0.4 μM for 1 h before the immunofluorescence staining for YAP and β-catenin. The images were taken using Cytation5 under 20× magnification. Scale bars = 100 μm (B) Cell populations with β-catenin nuclear translocation in A were measured using BioTek Gen5 Software, and the results were generated using GraphPad Prism. Data represent mean ± SD. n = 3 wells per condition. Statistical significance was assessed using one-way ANOVA followed by the Tukey post hoc test. ns: not significant, ∗ p < 0.05, ∗∗ p < 0.01, and ∗∗∗∗ p < 0.0001.

Journal: iScience

Article Title: A stiffness-gated YAP-β-catenin axis orchestrates AXIN2 expression in metastatic breast cancer

doi: 10.1016/j.isci.2025.114405

Figure Lengend Snippet: Myosin II activity facilitates YAP and β-catenin nuclear translocation, while intact actin filaments restrict β-catenin nuclear localization (A) MDA-MB-231 cells were transfected with non-targeting siRNA (control), siRNA against YAP (siYAP). Control or siYAP cells were treated with Blebbistatin at a concentration of 50 μM for 30 min or with LatA at a concentration of 0.4 μM for 1 h before the immunofluorescence staining for YAP and β-catenin. The images were taken using Cytation5 under 20× magnification. Scale bars = 100 μm (B) Cell populations with β-catenin nuclear translocation in A were measured using BioTek Gen5 Software, and the results were generated using GraphPad Prism. Data represent mean ± SD. n = 3 wells per condition. Statistical significance was assessed using one-way ANOVA followed by the Tukey post hoc test. ns: not significant, ∗ p < 0.05, ∗∗ p < 0.01, and ∗∗∗∗ p < 0.0001.

Article Snippet: MDA-MB-231 human breast cancer cell line , ATCC , HTB-26; RRID: CVCL_0062.

Techniques: Activity Assay, Translocation Assay, Transfection, Control, Concentration Assay, Immunofluorescence, Staining, Software, Generated

Matrix stiffness modulates total and phosphorylated β-catenin levels in breast cancer cells (A) Representative Western blots of total β-catenin and phosphorylated β-catenin at S33/S37/T41 and S675 in MCF-7 and MDA-MB-231 cells cultured on soft (2 kPa) and stiff (32 kPa) substrates. (B) Quantification of protein levels normalized to loading controls (mean ± SD). Top row: Total β-catenin expression showed no significant differences across stiffness in MCF-7 cells but was significantly increased in MDA-MB-231 cells cultured on stiff (32 kPa) compared to soft (2 kPa) substrates (∗ p < 0.05). Middle row: Phosphorylation of β-catenin at S33/S37/T41 (associated with degradation) was significantly higher in MDA-MB-231 cells than in MCF-7 cells at both stiffness levels (∗∗ p < 0.01), indicating enhanced β-catenin degradation in the metastatic subtype. However, substrate stiffness had no significant effect on p-β-catenin (S33/S37/T41) within either cell line. Bottom row: Phosphorylation at S675 (associated with nuclear translocation) was significantly higher in MCF-7 cells than in MDA-MB-231 cells at both stiffness levels ∗∗∗ p < 0.001). Data represent mean ± SD. n = 3 wells per condition. Statistical significance was assessed using two-tailed t-tests. ns: not significant; ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001.

Journal: iScience

Article Title: A stiffness-gated YAP-β-catenin axis orchestrates AXIN2 expression in metastatic breast cancer

doi: 10.1016/j.isci.2025.114405

Figure Lengend Snippet: Matrix stiffness modulates total and phosphorylated β-catenin levels in breast cancer cells (A) Representative Western blots of total β-catenin and phosphorylated β-catenin at S33/S37/T41 and S675 in MCF-7 and MDA-MB-231 cells cultured on soft (2 kPa) and stiff (32 kPa) substrates. (B) Quantification of protein levels normalized to loading controls (mean ± SD). Top row: Total β-catenin expression showed no significant differences across stiffness in MCF-7 cells but was significantly increased in MDA-MB-231 cells cultured on stiff (32 kPa) compared to soft (2 kPa) substrates (∗ p < 0.05). Middle row: Phosphorylation of β-catenin at S33/S37/T41 (associated with degradation) was significantly higher in MDA-MB-231 cells than in MCF-7 cells at both stiffness levels (∗∗ p < 0.01), indicating enhanced β-catenin degradation in the metastatic subtype. However, substrate stiffness had no significant effect on p-β-catenin (S33/S37/T41) within either cell line. Bottom row: Phosphorylation at S675 (associated with nuclear translocation) was significantly higher in MCF-7 cells than in MDA-MB-231 cells at both stiffness levels ∗∗∗ p < 0.001). Data represent mean ± SD. n = 3 wells per condition. Statistical significance was assessed using two-tailed t-tests. ns: not significant; ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001.

Article Snippet: MDA-MB-231 human breast cancer cell line , ATCC , HTB-26; RRID: CVCL_0062.

Techniques: Western Blot, Cell Culture, Expressing, Phospho-proteomics, Translocation Assay, Two Tailed Test

3D spheroid model reveals the stiffness-dependent regulation of YAP and β-catenin localization (A) Bright-field images of MDA-MB-231 spheroids grown in 3D Matrigel at 2, 4, and 8 mg/mL for 48–96 h post-siRNA transfection. Scale bars = 500 μm. (B) Quantification of spheroid spreading area shows no significant changes following YAP knockdown. Data represent mean ± SD. n = 4 wells per condition. Statistical significance was assessed using two-way ANOVA followed by the Tukey post hoc test. ns: not significant. (C) Cell viability measured by absorbance indicates a significant reduction in proliferation at all stiffness levels upon YAP knockdown. Data represent mean ± SD. n = 3 wells per condition. Statistical significance was assessed using two-way ANOVA followed by the Tukey post hoc test. ∗ p < 0.05, ∗∗ p < 0.01. (D) Violin plots show reduced YAP expression and β-catenin nuclear-to-cytoplasmic ratio in siYAP-treated spheroids. The effect on β-catenin localization is stiffness-dependent, observed at 2 and 4 mg/mL but not at 8 mg/mL. Data represent mean ± SD. n = individual cells pooled from 15 images (5 fields × 3 wells) per condition. Statistical significance was assessed using one-way ANOVA followed by the Tukey post hoc test. ns: not significant, ∗ p < 0.05, ∗∗∗∗ p < 0.0001.

Journal: iScience

Article Title: A stiffness-gated YAP-β-catenin axis orchestrates AXIN2 expression in metastatic breast cancer

doi: 10.1016/j.isci.2025.114405

Figure Lengend Snippet: 3D spheroid model reveals the stiffness-dependent regulation of YAP and β-catenin localization (A) Bright-field images of MDA-MB-231 spheroids grown in 3D Matrigel at 2, 4, and 8 mg/mL for 48–96 h post-siRNA transfection. Scale bars = 500 μm. (B) Quantification of spheroid spreading area shows no significant changes following YAP knockdown. Data represent mean ± SD. n = 4 wells per condition. Statistical significance was assessed using two-way ANOVA followed by the Tukey post hoc test. ns: not significant. (C) Cell viability measured by absorbance indicates a significant reduction in proliferation at all stiffness levels upon YAP knockdown. Data represent mean ± SD. n = 3 wells per condition. Statistical significance was assessed using two-way ANOVA followed by the Tukey post hoc test. ∗ p < 0.05, ∗∗ p < 0.01. (D) Violin plots show reduced YAP expression and β-catenin nuclear-to-cytoplasmic ratio in siYAP-treated spheroids. The effect on β-catenin localization is stiffness-dependent, observed at 2 and 4 mg/mL but not at 8 mg/mL. Data represent mean ± SD. n = individual cells pooled from 15 images (5 fields × 3 wells) per condition. Statistical significance was assessed using one-way ANOVA followed by the Tukey post hoc test. ns: not significant, ∗ p < 0.05, ∗∗∗∗ p < 0.0001.

Article Snippet: MDA-MB-231 human breast cancer cell line , ATCC , HTB-26; RRID: CVCL_0062.

Techniques: Transfection, Knockdown, Expressing

YAP knockdown selectively downregulates AXIN2 expression on soft substrates in metastatic breast cancer cells (A) Capillary-based immunoblot and quantification confirming efficient YAP knockdown in MDA-MB-231 cells cultured on soft (2 kPa) and stiff (32 kPa) substrates for 48 h. YAP protein levels were significantly reduced in siYAP-treated cells compared to control (∗∗∗∗ p < 0.0001). Data represent mean ± SD. n = 3 wells per condition. Statistical significance was assessed using two-way ANOVA followed by the Tukey post hoc test. (B) Immunoblot and quantification of CCND1 and AXIN2 protein expression in MDA-MB-231 and MCF-7 cells following YAP knockdown. In MDA-MB-231 cells, YAP depletion did not significantly affect CCND1 expression at either stiffness, while AXIN2 levels were significantly decreased under 2 kPa (∗ p < 0.05) but not 32 kPa. In MCF-7 cells, CCND1 expression was significantly elevated on stiff substrate compared to soft (∗ p < 0.05), independent of YAP knockdown. AXIN2 expression in MCF-7 cells was not significantly affected by YAP knockdown at either stiffness. Data represent mean ± SD. n = 3 wells per condition. Statistical significance was assessed using two-way ANOVA followed by the Tukey post hoc test. ns: not significant.

Journal: iScience

Article Title: A stiffness-gated YAP-β-catenin axis orchestrates AXIN2 expression in metastatic breast cancer

doi: 10.1016/j.isci.2025.114405

Figure Lengend Snippet: YAP knockdown selectively downregulates AXIN2 expression on soft substrates in metastatic breast cancer cells (A) Capillary-based immunoblot and quantification confirming efficient YAP knockdown in MDA-MB-231 cells cultured on soft (2 kPa) and stiff (32 kPa) substrates for 48 h. YAP protein levels were significantly reduced in siYAP-treated cells compared to control (∗∗∗∗ p < 0.0001). Data represent mean ± SD. n = 3 wells per condition. Statistical significance was assessed using two-way ANOVA followed by the Tukey post hoc test. (B) Immunoblot and quantification of CCND1 and AXIN2 protein expression in MDA-MB-231 and MCF-7 cells following YAP knockdown. In MDA-MB-231 cells, YAP depletion did not significantly affect CCND1 expression at either stiffness, while AXIN2 levels were significantly decreased under 2 kPa (∗ p < 0.05) but not 32 kPa. In MCF-7 cells, CCND1 expression was significantly elevated on stiff substrate compared to soft (∗ p < 0.05), independent of YAP knockdown. AXIN2 expression in MCF-7 cells was not significantly affected by YAP knockdown at either stiffness. Data represent mean ± SD. n = 3 wells per condition. Statistical significance was assessed using two-way ANOVA followed by the Tukey post hoc test. ns: not significant.

Article Snippet: MDA-MB-231 human breast cancer cell line , ATCC , HTB-26; RRID: CVCL_0062.

Techniques: Knockdown, Expressing, Western Blot, Cell Culture, Control

β-catenin knockdown reduces CTGF mRNA but not protein expression, and does not affect CCND1 expression (A) qPCR analysis showing efficient CTNNB1 knockdown and the CCND1 and CTGF transcript levels in MDA-MB-231 and MCF-7 cells. Data represent mean ± SD. n = 3 wells per condition. Statistical significance was assessed using two-way ANOVA followed by the Tukey post hoc test. ns: not significant, ∗∗ p < 0.01, ∗∗∗ p < 0.001, ∗∗∗∗ p < 0.0001. (B) Western blots and quantification confirm reduced β-catenin protein following siRNA treatment, but no significant change in CCND1 or CTGF protein expression. These results suggest that β-catenin alone is insufficient to regulate YAP target genes in the absence of YAP. Data represent mean ± SD. n = 3 wells per condition. Statistical significance was assessed using two-way ANOVA followed by the Tukey post hoc test. ns: not significant, ∗∗∗∗ p < 0.0001.

Journal: iScience

Article Title: A stiffness-gated YAP-β-catenin axis orchestrates AXIN2 expression in metastatic breast cancer

doi: 10.1016/j.isci.2025.114405

Figure Lengend Snippet: β-catenin knockdown reduces CTGF mRNA but not protein expression, and does not affect CCND1 expression (A) qPCR analysis showing efficient CTNNB1 knockdown and the CCND1 and CTGF transcript levels in MDA-MB-231 and MCF-7 cells. Data represent mean ± SD. n = 3 wells per condition. Statistical significance was assessed using two-way ANOVA followed by the Tukey post hoc test. ns: not significant, ∗∗ p < 0.01, ∗∗∗ p < 0.001, ∗∗∗∗ p < 0.0001. (B) Western blots and quantification confirm reduced β-catenin protein following siRNA treatment, but no significant change in CCND1 or CTGF protein expression. These results suggest that β-catenin alone is insufficient to regulate YAP target genes in the absence of YAP. Data represent mean ± SD. n = 3 wells per condition. Statistical significance was assessed using two-way ANOVA followed by the Tukey post hoc test. ns: not significant, ∗∗∗∗ p < 0.0001.

Article Snippet: MDA-MB-231 human breast cancer cell line , ATCC , HTB-26; RRID: CVCL_0062.

Techniques: Knockdown, Expressing, Western Blot

The miR-200:ZEB axis is prominent among putative microRNA: target gene pairs A - D Strength of miRNA target prediction (TargetScan score; more negative = more strongly predicted) and correlation of miRNA: putative target expression (REC score) across the CCLE or breast cancer TCGA. Transcription factors (foreground) are represented as red dots, all other genes as grey dots. Boxed section is enlarged in B, D. The 550 highest confidence miRNAs are included in this analysis E - F Correlation of ZEB and miR-200c expression across CCLE (E) and TCGA (breast cancer) (F) G Ranking of correlated expression from 10 cancer types across TCGA between ZEB1 (blue) or ZEB2 (red) and miR-200c. Genes are ranked from most to least correlated with horizontal lines indicating the ranked position of correlation with miR-200c relative to all other genes. The position of zero correlation (grey bar) is indicated H Ranked targeting strength of all putative miR-200c target genes as predicted by multiple algorithms. Number of targets predicted by each algorithm are shown at the bottom I Targetscan score and log fold change of all genes 3-days post miR-200c transfection in MDA-MB-231 cells. Transcription factors are again indicated by red dots. All other genes are represented as grey dots. The specific position of ZEB1 and ZEB2 are indicated. Dotted line indicates the 5th percentiles of the most strongly predicted targets and of genes downregulated in response to miR-200c

Journal: Cell Communication and Signaling : CCS

Article Title: Strongly regulated transcription factors exert an outsized influence in microRNA-regulated networks

doi: 10.1186/s12964-025-02626-w

Figure Lengend Snippet: The miR-200:ZEB axis is prominent among putative microRNA: target gene pairs A - D Strength of miRNA target prediction (TargetScan score; more negative = more strongly predicted) and correlation of miRNA: putative target expression (REC score) across the CCLE or breast cancer TCGA. Transcription factors (foreground) are represented as red dots, all other genes as grey dots. Boxed section is enlarged in B, D. The 550 highest confidence miRNAs are included in this analysis E - F Correlation of ZEB and miR-200c expression across CCLE (E) and TCGA (breast cancer) (F) G Ranking of correlated expression from 10 cancer types across TCGA between ZEB1 (blue) or ZEB2 (red) and miR-200c. Genes are ranked from most to least correlated with horizontal lines indicating the ranked position of correlation with miR-200c relative to all other genes. The position of zero correlation (grey bar) is indicated H Ranked targeting strength of all putative miR-200c target genes as predicted by multiple algorithms. Number of targets predicted by each algorithm are shown at the bottom I Targetscan score and log fold change of all genes 3-days post miR-200c transfection in MDA-MB-231 cells. Transcription factors are again indicated by red dots. All other genes are represented as grey dots. The specific position of ZEB1 and ZEB2 are indicated. Dotted line indicates the 5th percentiles of the most strongly predicted targets and of genes downregulated in response to miR-200c

Article Snippet: The MDA-MB-231 (ATCC: HTB-26) immortalised human breast cancer cell line was obtained from American Type Culture Collection (ATCC, Rockville, MD) and used to generate ZEB-expressing stable cell lines.

Techniques: Expressing, Transfection

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