Journal: Bioactive Materials

Article Title: Harnessing the power of physicochemical material property screening to direct breast epithelial and breast cancer cells

doi: 10.1016/j.bioactmat.2025.04.003

Figure Lengend Snippet: DOG screening overview. a , Screening overview showing the heatmaps (average of n = 3) of the cell proliferation at 24 h and 72 h for both the MCF10a and MCF7 cells quantified as Ki-67 positive nuclei. The heatmaps corresponding to the cell density and cell area can be found in the supplementary figures. b , Scatter plots showing all data points from one screening (4 DOGs) ranked from low to high. Tissue culture polystyrene (TCP) was used as a control. The 10 % lowest and 10 % highest scoring datapoints are shown on the side and are shown to be significantly different (p < 0.0001). Annotation of black dot with “T” or “L” corresponds to the representative fluorescence images of “Top” and “Low” hits as shown in .

Article Snippet: Normal breast epithelial cells (MCF10a, Aldrich) were maintained in MCF10a Complete Medium (EP-ML-0525, Elabscience) containing 5 % horse serum, 20 ng mL −1 epidermal growth factor (EGF), 0.5 μg/mL hydrocortisone, 10 μg/mL insulin, 1 % non-essential amino acid solution (NEAA), and 1 % penicillin/streptomycin.

Techniques: Control, Fluorescence

Journal: Bioactive Materials

Article Title: Harnessing the power of physicochemical material property screening to direct breast epithelial and breast cancer cells

doi: 10.1016/j.bioactmat.2025.04.003

Figure Lengend Snippet: Top- and low-hit Ki-67-expressing proliferative cells after 72 h of cell culture . Cells were stained for Ki-67 (red), phalloidin (green), and nuclei (blue). Scale bar is 100 μm. White arrows show examples of Ki-67 positive nuclei. The underlying material properties of each image are: MCF10a top hit: S-W 51° 251 MPa, MCF10a low hit: T-S|W 3.8 μm 82° 47 MPa, MCF7 top hit: S-W 63° 42 MPa, MCF7 low hit: T-SW 6 μm 68° 63 MPa.

Article Snippet: Normal breast epithelial cells (MCF10a, Aldrich) were maintained in MCF10a Complete Medium (EP-ML-0525, Elabscience) containing 5 % horse serum, 20 ng mL −1 epidermal growth factor (EGF), 0.5 μg/mL hydrocortisone, 10 μg/mL insulin, 1 % non-essential amino acid solution (NEAA), and 1 % penicillin/streptomycin.

Techniques: Expressing, Cell Culture, Staining

Journal: Bioactive Materials

Article Title: Harnessing the power of physicochemical material property screening to direct breast epithelial and breast cancer cells

doi: 10.1016/j.bioactmat.2025.04.003

Figure Lengend Snippet: Scatter plots comparing the effect of each individual physicochemical parameter of the DOG between MCF10a and MCF7 cells. The material parameters (topography, wettability, and stiffness on the same data points, shown with the corresponding color scale) were shown on the biological output (cell proliferation, % Ki-67 positive cells) for the MCF10a vs. the MCF7 cells. Similar plots showing the scatter plots for cell density and cell area can be found in .

Article Snippet: Normal breast epithelial cells (MCF10a, Aldrich) were maintained in MCF10a Complete Medium (EP-ML-0525, Elabscience) containing 5 % horse serum, 20 ng mL −1 epidermal growth factor (EGF), 0.5 μg/mL hydrocortisone, 10 μg/mL insulin, 1 % non-essential amino acid solution (NEAA), and 1 % penicillin/streptomycin.

Techniques:

Journal: Bioactive Materials

Article Title: Harnessing the power of physicochemical material property screening to direct breast epithelial and breast cancer cells

doi: 10.1016/j.bioactmat.2025.04.003

Figure Lengend Snippet: ROIs were chosen that lead to a higher proliferation after 72 h for MCF10a than MCF7 cells. a , Regions of interest (ROIs) are chosen as points furthest away from the dashed line. In orange and blue, points are indicated that show an individual statistical difference (P < 0.05) when comparing the proliferation percentage of MCF7 vs. MCF10a. Orange or blue dots indicate an increase for either MCF7 or MCF10a proliferation, respectively. b , Points from a were grouped and show a significant different in both MCF10a and MCF7 Ki-67 positive cells percentage (p < 0.0001). c , Individual DOG values for each ROI chosen in a are shown. Corresponding fluorescence images for each ROI can be found in .

Article Snippet: Normal breast epithelial cells (MCF10a, Aldrich) were maintained in MCF10a Complete Medium (EP-ML-0525, Elabscience) containing 5 % horse serum, 20 ng mL −1 epidermal growth factor (EGF), 0.5 μg/mL hydrocortisone, 10 μg/mL insulin, 1 % non-essential amino acid solution (NEAA), and 1 % penicillin/streptomycin.

Techniques: Fluorescence

Journal: Bioactive Materials

Article Title: Harnessing the power of physicochemical material property screening to direct breast epithelial and breast cancer cells

doi: 10.1016/j.bioactmat.2025.04.003

Figure Lengend Snippet: ROIs were chosen from the screening as “positive” when MCF10a Ki-67 % > MCF7 Ki-67 % (labelled as Pos1, Pos2, Pos3, shown in blue) and “negative” when MCF7 Ki-67 % > MCF10a Ki-67 % (labelled as Neg1, Neg2, Neg3, shown in orange). a , Cell experiments were repeated on translational (labelled as Trans) substrates with those specific material properties and compared to the proliferation (% Ki-67 positive cells after 72 h of cell culture) rates from the screening (Screen). b , Cell density after 72 h was compared between the screening and translation findings. c , Fluorescence images of two example ROIs, Pos1 and Neg1. Cells were stained for Ki-67 (red), phalloidin (green), and DAPI (blue). Corresponding fluorescence images for the other ROIs can be found in . The scale bar is 100 μm.

Article Snippet: Normal breast epithelial cells (MCF10a, Aldrich) were maintained in MCF10a Complete Medium (EP-ML-0525, Elabscience) containing 5 % horse serum, 20 ng mL −1 epidermal growth factor (EGF), 0.5 μg/mL hydrocortisone, 10 μg/mL insulin, 1 % non-essential amino acid solution (NEAA), and 1 % penicillin/streptomycin.

Techniques: Cell Culture, Fluorescence, Staining

Journal: Bioactive Materials

Article Title: Harnessing the power of physicochemical material property screening to direct breast epithelial and breast cancer cells

doi: 10.1016/j.bioactmat.2025.04.003

Figure Lengend Snippet: Proliferation quantified as % Ki-67 positive cells after 72 h for both MCF7 and MCF10a cells compared between the translational cell experiments on translated ROIs for both low (shown in blue, 1000 cells cm −2 ) and high (shown in orange, 5000 cells cm −2 ) seeding density. a , Individual values for each ROI compared between low and high seeding density. b , All low and high seeding density datapoints from a were grouped and compared for statistical difference (n.s.: p > 0.05, ∗: p < 0.05).

Article Snippet: Normal breast epithelial cells (MCF10a, Aldrich) were maintained in MCF10a Complete Medium (EP-ML-0525, Elabscience) containing 5 % horse serum, 20 ng mL −1 epidermal growth factor (EGF), 0.5 μg/mL hydrocortisone, 10 μg/mL insulin, 1 % non-essential amino acid solution (NEAA), and 1 % penicillin/streptomycin.

Techniques:

Journal: Proceedings of the National Academy of Sciences of the United States of America

Article Title: Control of cyclooxygenase-2 expression and tumorigenesis by endogenous 5-methoxytryptophan

doi: 10.1073/pnas.1209919109

Figure Lengend Snippet: Suppression of cancer cell COX-2 expression by soluble factors from fibroblasts. (A) A549 lung cancer cells (106 cells) were cocultured with HsFb at various cell densities in a Boyden chamber for 24 h. A549 cells were removed and treated with PMA (100 nM) for 4 h. COX-2 proteins in the cell lysates were analyzed by Western blotting. (B) Conditioned medium (CM) was collected from Hs68 fibroblasts (HsFb) or A549 cells cultured in medium containing 2.5% FBS for 24 h. (Left) HsFb CM was added to washed A549 cells. (Right) A549 CM was added to washed serum-starved Hs68 fibroblasts. Cells were treated with PMA for 4 h, and COX-2 proteins were analyzed by Western blotting. Error bars denote mean ± SEM (n = 3). (C) CMF2 fractions prepared from HsFb or A549 CM were added to washed A549, HT29, or serum-starved HsFb, and PMA-induced COX-2 was analyzed. CMF2 of HsFb inhibited COX-2 expression whereas CMF2 of A549 had no effect. Two independent experiments were performed with similar results. (D) Representative mass spectra (m/z 0–300) of CMF2 prepared from HsFb or A549. Control denotes F2 fraction of the basal DMEM containing 2.5% FBS. Metabolomic analysis of CMF2 was performed using UPLC–QTof mass spectrometry. (E) Comparison of relative intensity of m/z 276.1 and m/z 262.1 between HsFb CMF2 and A549 CMF2. Error bars denote mean ± SEM (n = 3). (F) Effect of cancer-associated fibroblasts on PMA-induced COX-2 expression in A549 cells. CM and CMF2 prepared from prostate cancer-associated fibroblasts (PcFb) inhibited COX-2 expression whereas neither CM nor CMF2 of breast cancer-associated fibroblasts (bCAF) did. (G and H) The metabolomic profile of (G) PcFb-CMF2 was similar to that of HsFb-CMF2, whereas the profile of (H) bCAF-CMF2 reveals diminished m/z 276.1, m/z 262.1, and other m/z peaks.

Article Snippet: Human cancer cell lines A549, HT-29, Hep3B, and MCF7 and MCF10A were from the Bioresource Collection and Research Center.

Techniques: Expressing, Western Blot, Cell Culture, Control, Mass Spectrometry, Comparison

Journal: Proceedings of the National Academy of Sciences of the United States of America

Article Title: Control of cyclooxygenase-2 expression and tumorigenesis by endogenous 5-methoxytryptophan

doi: 10.1073/pnas.1209919109

Figure Lengend Snippet: Tryptophan hydroxylase (TPH-1)-derived 5-HTP suppresses COX-2 expression. (A) TPH-1 proteins in HsFb vs. A549 cells were analyzed by Western blotting. The error bars refer to mean ± SEM (n = 3). (B) TPH-1 protein levels in siRNA-transfected HsFb vs. control scRNA-transfected HsFb. “Basal” denotes untransfected cells. (C) Analysis of COX-2 proteins in HsFb transfected with TPH-1 siRNA or scRNA. COX-2 was increased in siRNA-treated cells, and COX-2 suppression was restored by 5-HTP (10 μM) or 5-MTP (10 μM) supplement. 5-MTP supplement completely inhibited COX-2 expression whereas 5-HTP partially suppressed COX-2 proteins. (D) HsFb were pretreated with 5-HTP followed by PMA for 4 h. COX-2 proteins were analyzed by Western blotting. (E) Metabolomic analysis of CMF2 prepared from CM of HsFb with or without siRNA or scRNA transfection.

Article Snippet: Human cancer cell lines A549, HT-29, Hep3B, and MCF7 and MCF10A were from the Bioresource Collection and Research Center.

Techniques: Derivative Assay, Expressing, Western Blot, Transfection, Control

Journal: Proceedings of the National Academy of Sciences of the United States of America

Article Title: Control of cyclooxygenase-2 expression and tumorigenesis by endogenous 5-methoxytryptophan

doi: 10.1073/pnas.1209919109

Figure Lengend Snippet: Endogenous and exogenously added 5-MTP inhibits COX-2 expression. (A) Pure 5-MTP at increasing concentrations was added to A549 treated with or without PMA for 4 h. COX-2 proteins were analyzed by Western blotting. (B) 5-MTP restored COX-2 suppression in TPH-1 siRNA-transfected HsFb in a concentration-dependent manner. (C) Addition of 5-MTP to A549 cells enhances m/z 276.1 and 262.1 peaks on mass spectrometry. (D) HsFbs were transfected with TPH-1 siRNA or scRNA. l-Tryptophan (Try) or 5-HTP was added to the transfected cells. Conditioned medium was collected, and 5-MTP concentration was measured by EIA. Each bar denotes mean ± SEM (n = 3). ns, not statistically significant. (E) Analysis of COX-2 proteins by Western blotting in HsFb transfected with HIOMT siRNA or a control scRNA. (Upper) Representative blot. (Lower) Quantitative analysis by densitometry. Error bars denote mean ± SEM (n = 3). Concentration of 5-MTP was 10 μM. (F) Measurement by EIA of 5-MTP in the CM of HsFb under the indicated treatment; 10 μM of 5-HTP or 5-MTP was added. Each error bar denotes mean ± SEM of three independent experiments done in triplicate. ns, not statistically significant.

Article Snippet: Human cancer cell lines A549, HT-29, Hep3B, and MCF7 and MCF10A were from the Bioresource Collection and Research Center.

Techniques: Expressing, Western Blot, Transfection, Concentration Assay, Mass Spectrometry, Control

Journal: Proceedings of the National Academy of Sciences of the United States of America

Article Title: Control of cyclooxygenase-2 expression and tumorigenesis by endogenous 5-methoxytryptophan

doi: 10.1073/pnas.1209919109

Figure Lengend Snippet: 5-MTP rescues A549 defects in 5-MTP synthesis and cell proliferation and migration. (A) Analysis of HIOMT proteins in A549 vs. HsFb. (Upper) Representative blot. (Lower) Densitometric analysis of three independent experiments. The error bars are mean ± SEM (*P < 0.01). (B) Measurement of 5-MTP level in A549 vs. HsFb CM. Error bars are mean ± SEM (n = 3). (C) Neither Try nor 5-HTP increased A549 5-MTP production, whereas addition of exogenous 5-MTP increased the 5-MTP level measured by EIA. The error bars are mean ± SEM (n = 3). (D) A549 cells labeled with MTT were treated with 5-MTP for 4 or 24 h. Each bar denotes mean ± SEM (n = 3). (E) A549 cells were pretreated with 5-MTP (10 μM) for 30 min followed by PMA (100 nM) for 6 or 24 h. (Upper panels) Representative cell migration. (Lower panels) Migrated cell counts expressed as percentage of basal controls. Error bars denote mean ± SEM (n = 3). (F) A549 cells were seeded on regular or matrigel-coated membrane and pretreated with 5-MTP for 30 min followed by PMA. (Upper) Representative cell migration and invasion. (Lower panels) Quantitative analysis of migration and invasion.

Article Snippet: Human cancer cell lines A549, HT-29, Hep3B, and MCF7 and MCF10A were from the Bioresource Collection and Research Center.

Techniques: Migration, Labeling, Membrane

Journal: Proceedings of the National Academy of Sciences of the United States of America

Article Title: Control of cyclooxygenase-2 expression and tumorigenesis by endogenous 5-methoxytryptophan

doi: 10.1073/pnas.1209919109

Figure Lengend Snippet: 5-MTP reduces cancer growth and metastasis in a murine tumor xenograft model. A549 (5 × 106 cells) were transfected with luciferase (A549-luc-c8) and injected s.c. into the flank of SCID-Beige mice. Ten mice each received i.p. injection of 5-MTP (100 mg/kg) or vehicle twice weekly. (A) Caliper measurement of the s.c. tumor volume periodically for 7 wk. *P < 0.05 and **P < 0.01. (B) Tumor growth was monitored by in vivo bioluminescent imaging (IVIS). The time courses of IVIS of two representative vehicle-treated and two representative 5-MTP–treated mice are illustrated. (C) 5-MTP reduces lung metastasis. Mice were euthanized and both lungs were removed. Nodules were counted. Each nodule was sectioned and stained with hematoxylin-eosine and examined under the microscope. 5-MTP pretreatment significantly attenuated lung metastasis. (D) Metabolic scheme illustrating 5-MTP biosynthesis in HsFb vs. A549 cancer cells. The dotted line denotes possible defects in the synthetic pathway.

Article Snippet: Human cancer cell lines A549, HT-29, Hep3B, and MCF7 and MCF10A were from the Bioresource Collection and Research Center.

Techniques: Transfection, Luciferase, Injection, In Vivo, Imaging, Staining, Microscopy

Journal: Oncology Letters

Article Title: Expression and anti-apoptotic function of TRAF4 in human breast cancer MCF-7 cells

doi: 10.3892/ol.2013.1703

Figure Lengend Snippet: Expression and location of TRAF4 in breast cancer cells. (A) Results of immunofluorescence staining showing TRAF4 (green fluorescence) localized in the cytoplasm and nuclei (scale bar, 20 μm). (B) MCF-10A, MCF-7 and MDA-MB-231 cells were lysed and the lysates were subjected to western blot analyses of TRAF4 and β-actin. Results showed that the total expression of TRAF4 in the MCF-10A cells was lower than in the MCF-7 and MDA-MB-231 cells ( ** P=0.002 and P=0.001, respectively). TRAF4, tumor necrosis factor receptor-associated factor 4; IDV, integated optical density value.

Article Snippet: The membranes for western blotting were incubated with mouse anti-human monoclonal antibodies against TRAF4 (1:1,000; BD Biosciences, Franklin Lakes, NJ, USA), NF-κB p65 (1:1,000; Beyotime Institute of Biotechnology, Haimen, China), lamin B1 (1:500; Santa Cruz Biotechnology, Inc., Santa Cruz, CA, USA) and β-actin (1:1,000; Santa Cruz Biotechnology Inc.), followed by horseradish peroxidase-conjugated secondary antibody (Santa Cruz Biotechnology Inc.).

Techniques: Expressing, Immunofluorescence, Staining, Fluorescence, Western Blot

Journal: Oncology Letters

Article Title: Expression and anti-apoptotic function of TRAF4 in human breast cancer MCF-7 cells

doi: 10.3892/ol.2013.1703

Figure Lengend Snippet: TRAF4 suppresses the activation of NF-κB in MCF-7 Cells. The MCF-7 cells were lysed and the lysates were subjected to western blot analyses of NF-κB, β-actin and lamin B1. Following TNF-α treatment (10 ng/ml; 15 min), the nuclear expression of NF-κB was significantly downregulated when TRAF4 was knocked down in the MCF-7 cells ( ** P=0.002). TRAF4, tumor necrosis factor receptor-associated factor 4; NF-κB, nuclear factor κB.

Article Snippet: The membranes for western blotting were incubated with mouse anti-human monoclonal antibodies against TRAF4 (1:1,000; BD Biosciences, Franklin Lakes, NJ, USA), NF-κB p65 (1:1,000; Beyotime Institute of Biotechnology, Haimen, China), lamin B1 (1:500; Santa Cruz Biotechnology, Inc., Santa Cruz, CA, USA) and β-actin (1:1,000; Santa Cruz Biotechnology Inc.), followed by horseradish peroxidase-conjugated secondary antibody (Santa Cruz Biotechnology Inc.).

Techniques: Activation Assay, Western Blot, Expressing

Journal: Oncology Letters

Article Title: Expression and anti-apoptotic function of TRAF4 in human breast cancer MCF-7 cells

doi: 10.3892/ol.2013.1703

Figure Lengend Snippet: Biological function of TRAF4 in MCF-7 cells. (A) MTT results showing that cell proliferation was significantly suppressed following the knockdown of TRAF4 ( * P=0.02). Results of flow cytometry showed that TRAF4 ablation resulted in (B) significant increase of G 1 phase cells ( ** P=0.009) and reduction of S phase cells ( * P=0.04) and (C) significant increase in the early apoptotic cells ( ** P=0.001). All comparisons were made between the groups of MCF-7 cells or cells transfected with negative control alone. TRAF4, tumor necrosis factor receptor-associated factor 4.

Article Snippet: The membranes for western blotting were incubated with mouse anti-human monoclonal antibodies against TRAF4 (1:1,000; BD Biosciences, Franklin Lakes, NJ, USA), NF-κB p65 (1:1,000; Beyotime Institute of Biotechnology, Haimen, China), lamin B1 (1:500; Santa Cruz Biotechnology, Inc., Santa Cruz, CA, USA) and β-actin (1:1,000; Santa Cruz Biotechnology Inc.), followed by horseradish peroxidase-conjugated secondary antibody (Santa Cruz Biotechnology Inc.).

Techniques: Flow Cytometry, Transfection, Negative Control

Journal: Journal of Oncology

Article Title: Eupalinolide O Induces Apoptosis in Human Triple-Negative Breast Cancer Cells via Modulating ROS Generation and Akt/p38 MAPK Signaling Pathway

doi: 10.1155/2022/8802453

Figure Lengend Snippet: EO suppressed the viability of human TNBC cells. The viability of TNBC cell lines MDA-MB-231 and MDA-MB-453 and normal epithelial cell line MCF 10A treated with different concentrations of EO for 24 h, 48 h, and 72 h measured by MTT. Unless otherwise specified, all results were presented as mean ± SD. n = 3. EO, Eupalinolide O; TNBC, Triple-negative breast cancer; MTT, 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide. # p < 0.05, and ## p < 0.01 vs. Control, in all figures.

Article Snippet: Human normal breast epithelial cell line MCF 10A and TNBC cell lines (MDA-MB-231 cells and MDA-MB-453 cells) were gifted by iCell Bioscience Inc (Shanghai, China).

Techniques: Control

Journal: Journal of Oncology

Article Title: Eupalinolide O Induces Apoptosis in Human Triple-Negative Breast Cancer Cells via Modulating ROS Generation and Akt/p38 MAPK Signaling Pathway

doi: 10.1155/2022/8802453

Figure Lengend Snippet: EO suppressed colony formation of human TNBC cells. The proliferation of MDA-MB-231, MDA-MB-453, and MCF 10A cells treated with different concentrations of EO detected by colony formation assay. n = 3.

Article Snippet: Human normal breast epithelial cell line MCF 10A and TNBC cell lines (MDA-MB-231 cells and MDA-MB-453 cells) were gifted by iCell Bioscience Inc (Shanghai, China).

Techniques: Colony Assay

Journal: Cell Communication and Signaling : CCS

Article Title: Interplay of the transcription factor MRTF-A and matrix stiffness controls mammary acinar structure and protrusion formation

doi: 10.1186/s12964-022-00977-2

Figure Lengend Snippet: Correlation of increasing matrix stiffness with protrusion formation of MCF10A acini. 3D cultures of MCF10A cells on the indicated Matrigel/Collagen I mixtures (100:0, 75:25, 50:50, 25:75, 0:100) were analysed by microscopy. A Phalloidin (green), Laminin V (red) and nuclei (DAPI, blue) staining of acinar structures at day 4 after seeding on matrices with increasing Collagen I percentages. Arrowheads indicate protrusions. The corresponding increase of Young's elastic modulus of the matrix is depicted below. B Diameters of individual acini (diamonds) measured at day 4, day 8 and day 14. Black horizontal lines indicate mean diameter and SEM (whiskers) of three biologically independent replicates. C Quantification of protrusion formation at day 4, day 8 and day 14. At least 11 acini were assessed each day, the percentage of protrusion-positive acini calculated, and the entire experiment repeated 3 times. D Calculated roundness index of individual acini (diamonds). Statistical significance compared to acini formed on pure Matrigel was determined using a one-way ANOVA and the means for each time point. * p ≤ 0.05, ** p ≤ 0.01, ***p ≤ 0.001. Error bars , SEM ( n = 3). Scale bars , 50 μm

Article Snippet: For 3D culture MCF10A growth medium with 2% horse serum and 2% Matrigel (Corning) was used.

Techniques: Microscopy, Staining

Journal: Cell Communication and Signaling : CCS

Article Title: Interplay of the transcription factor MRTF-A and matrix stiffness controls mammary acinar structure and protrusion formation

doi: 10.1186/s12964-022-00977-2

Figure Lengend Snippet: Acinar protrusions are independent of composition but depend on stiffness of matrix. A , B , C Matrigel (MG) or collagen I (COL1) gels were thinly coated with collagen IV (COL4) or left uncoated (wo COL4) as indicated. Subsequently MCF10A cells were seeded on top and acinar diameter A and protrusion formation B was analysed 4 days post seeding. C Representative micrographs of acini stained with laminin V (red), actin filaments with phalloidin (green) and cell nuclei with DAPI (blue) on day 4 of acini morphogenesis. Arrowheads indicate protrusions. D , E , F MCF10A cells were seeded on pre-cast acrylamide gels with defined stiffness of 0.2 kPa and 4 kPa coated with Matrigel, respectively. Acini diameter D and average percentage of protruding acini ( E ) were determined 4, 8 and 14 days post seeding as before. F Representative micrographs of acini on day 4 after seeding on the matrix indicated. Statistical significance was determined by ANOVA using the means of 3 independent experiments, each analysing ≥ 22 acini per condition. G , H Relative mRNA expression of ACTA2 G and VIM H normalised to ALAS and GAPDH. Statistical significance according to an unpaired Student’s t -test ( n = 4). * p ≤ 0.05, ** p ≤ 0.01. Error bars , SEM. Scale bars , 50 μm

Article Snippet: For 3D culture MCF10A growth medium with 2% horse serum and 2% Matrigel (Corning) was used.

Techniques: Staining, Expressing

Journal: Cell Communication and Signaling : CCS

Article Title: Interplay of the transcription factor MRTF-A and matrix stiffness controls mammary acinar structure and protrusion formation

doi: 10.1186/s12964-022-00977-2

Figure Lengend Snippet: Implication of the MRTF-SRF pathway in stiffness-dependent acini morphogenesis. A Relative MRTF/SRF luciferase reporter activity in transiently transfected MCF10A seeded on Matrigel (MG) or collagen I (COLI) and analysed at day 3 of acini morphogenesis ( n = 4). B , C Effects of the Rho/MRTF/SRF pathway inhibitor CCG203971 (20 μM, day 4–8) on acini diameter and average percentage of protrusion-positive acini at day 8. Eleven or more acini were analysed per condition in 3 biologically independent replicates. B Individually measured acini diameter (diamond) and mean ± SEM (horizontal black line ± whiskers). C Percentage of protrusion formation. Significance was tested using an unpaired Student’s t-test. * p ≤ 0.05. Error bars , SEM

Article Snippet: For 3D culture MCF10A growth medium with 2% horse serum and 2% Matrigel (Corning) was used.

Techniques: Luciferase, Activity Assay, Transfection