breast cancer cell lines mda mb 231  (ATCC)

Journal: Bioactive Materials

Article Title: Ribbon-shaped microgels as bioinks for 3D bioprinting of anisotropic tissue structures

doi: 10.1016/j.bioactmat.2025.12.040

Figure Lengend Snippet: μRB bioinks support multicellular patterning to model breast cancer-bone invasion at tissue interface. (A) Schematic of experimental design: bioprinting of MSCs and osteogenic differentiation to derive bone grid, followed by injection of breast cancer bioink, and monitoring invasion over time using confocal microscopy. (B) Confocal images of MSCs (red) after printing (Scale bar = 1 mm). (C) Confocal images of scaffold sections containing CellTracker-labeled MSCs (red) after 28 days of osteogenic differentiation and GFP + MDA-MB-231 cells extruded into the open pores of the grids (green) (Scale bar = 200 μm). (D) Confocal images of patterned MSC-derived bone (red) with MDA-MB-231 and MCF-7 breast cancer cells (green) after 14 days of co-culture (Scale bar = 1 mm). (E) Quantification of breast cancer cell invasion: percentage that remain in open pores vs. invading into the MSC-bone compartment (n = 5 per group). Values are reported as mean ± S.D. and p-values were determined by two-way analysis of variance (ANOVA) with Tukey's multiple comparisons test; ∗∗p ≤ 0.01, ∗∗∗p ≤ 0.005, ∗∗∗∗p ≤ 0.001.

Article Snippet: Breast cancer cell lines MDA-MB-231 (ATCC) and MCF-7 (ATCC) were lentivirus transduced to express GFP and cultured in DMEM media (4.5 g L −1 glucose) supplemented with 10 % (v/v) fetal bovine serum and 1 % (v/v) penicillin-streptomycin.

Techniques: Injection, Confocal Microscopy, Labeling, Derivative Assay, Co-Culture Assay

Journal: Free radical biology & medicine

Article Title: Redox regulation of cell migration via Nischarin S-glutathionylation

doi: 10.1016/j.freeradbiomed.2025.11.013

Figure Lengend Snippet: ( a ) NISCH structure, domains, and selected cysteines, including C185 (stick model). The structure was retrieved from the AlphaFold database. PX, LRR, CC, and ITGBD were found in UniProt. TED, PH-like, and PH are found in the AlphaFold and Pfam databases. ( b ) The accessible surface area (ASA) of cysteines in NISCH. ( c ) C185 and its neighboring residues in NISCH orthologs. ( d ) The outline of the clickable glutathione approach. Upon incubating azido-Ala, cells expressing a glutathione synthetase mutant (GS M4) synthesize and produce clickable glutathione (N 3 -GSH), which forms glutathionylation with proteins. ( e-f ) Glutathionylation of NISCH WT and cysteine mutants in HEK293/GS M4 cells in response to hydrogen peroxide (H 2 O 2 ). H 2 O 2 was incubated for 15 min (n = 3). ( g-h ) Glutathionylation of NISCH WT and C185S in MDA-MB-231 cells expressing GS M4 in low glucose (LG) compared to high glucose (HG) conditions. Cells were incubated in LG or HG for 16 h (n = 3–4). The lysates were subjected to click chemistry using biotin-alkyne and analyzed by Western blot using FLAG antibody before (input) and after (eluted) pull-down with streptavidin-agarose. Data represent the mean ± SD. The statistical difference was analyzed by one-way ANOVA and Tukey’s post-hoc test ( e , g - h ), where *p < 0.03, **p < 0.002, ***p < 0.0002, ****p < 0.0001.

Article Snippet: Human embryonic kidney 293 (HEK293) cell line stably expressing GS M4 (HEK293-GS M4) [ ], breast cancer cell line MDA-MB-231 (ATCC, HTB-26), and breast cancer cell line MCF-7 (ATCC, HTB-22) were maintained in Dulbecco’s modified Eagle’s medium (DMEM) supplemented with 10 % fetal bovine serum (Hyclone, Cytiva), penicillin (100 units/mL) and streptomycin (100 μg/mL) (Pen-Strep, Invitrogen) at 37°C in a 5 % CO 2 environment.

Techniques: Expressing, Mutagenesis, Incubation, Western Blot

Journal: Free radical biology & medicine

Article Title: Redox regulation of cell migration via Nischarin S-glutathionylation

doi: 10.1016/j.freeradbiomed.2025.11.013

Figure Lengend Snippet: ( a ) NISCH domain and its interacting partners. NISCH domains interacting with protein partners were drawn in lines in different colors. ( b-d ) Analysis of NISCH-protein interactions and Rac1 downstream pathway in MDA-MB-231 cells. MDA-MB-231 cells expressing NISCH WT or C185S were incubated in low or high glucose (LG or HG). ( b ) NISCH glutathionylation dissociates NISCH from Rac1 and PAK1. After incubating cells in LG or HG for 16 h, the co-immunoprecipitation was used to examine the binding of Rac1, PAK1, or integrin α5 (n = 2–3). ( c ) Rac1 activation analysis after NISCH C185 glutathionylation. After incubating cells for 16 h, the GST-fused p21-binding domain (PBD) derived from PAK1 (GST-PAK1-PBD) was used to pull down the Rac1-GTP form, which was then analyzed by Western blot (n = 2). ( d ) The co-localization analysis of NISCH and Rac1. After 16 h, cells were fixed and analyzed by antibodies to FLAG (green) and Rac1/Cdc42 (red) (n = 10 images). The merged images were analyzed to examine the relative displacement of NISCH and Rac1 in the cell periphery at higher magnification (box). Data represent the mean ± SD. The statistical difference was analyzed by one-way ANOVA and Tukey’s post-hoc test ( b-d ), where *p < 0.03, **p < 0.002, ***p < 0.0002, ****p < 0.0001.

Article Snippet: Human embryonic kidney 293 (HEK293) cell line stably expressing GS M4 (HEK293-GS M4) [ ], breast cancer cell line MDA-MB-231 (ATCC, HTB-26), and breast cancer cell line MCF-7 (ATCC, HTB-22) were maintained in Dulbecco’s modified Eagle’s medium (DMEM) supplemented with 10 % fetal bovine serum (Hyclone, Cytiva), penicillin (100 units/mL) and streptomycin (100 μg/mL) (Pen-Strep, Invitrogen) at 37°C in a 5 % CO 2 environment.

Techniques: Expressing, Incubation, Immunoprecipitation, Binding Assay, Activation Assay, Derivative Assay, Western Blot

Journal: Free radical biology & medicine

Article Title: Redox regulation of cell migration via Nischarin S-glutathionylation

doi: 10.1016/j.freeradbiomed.2025.11.013

Figure Lengend Snippet: MDA-MB-231 cells expressing NISCH WT or C185S were incubated in low or high glucose (LG or HG) for 16 h. ( a ) Phosphorylation levels of PAK1, ERK1/2, and MEK1/2 (n = 3). ( b ) Phosphorylation levels of LIMK1 and cofilin (n = 3). ( c ) Paxillin phosphorylation level (n = 3). ( d ) A model for cell migration induced by NISCH glutathionylation. In a non-stressed condition, NISCH binds to its protein partners, including Rac1 and PAK1, thus suppressing Rac1-and PAK1-mediated cell migration. NISCH may bind and retain Rac1 and integrin-α5 in the cytosol or endosome. However, upon ROS production, NISCH is glutathionylated at C185, dissociating Rac1 and PAK1, but not integrin-α5. The activated PAK1 activates its downstream LIMK1 and inhibits cofilin through phosphorylation, thereby increasing actin dynamics and polymerization. The dissociated Rac1 increases its localization to the membrane, where it activates actin filament branching, induces lamellipodia and membrane ruffles, and increases cell migration. Data represent the mean ± SD. The statistical difference was analyzed by one-way ANOVA and Tukey’s post-hoc test ( a – c ), where *p < 0.03, **p < 0.002, ***p < 0.0002, ****p < 0.0001.

Article Snippet: Human embryonic kidney 293 (HEK293) cell line stably expressing GS M4 (HEK293-GS M4) [ ], breast cancer cell line MDA-MB-231 (ATCC, HTB-26), and breast cancer cell line MCF-7 (ATCC, HTB-22) were maintained in Dulbecco’s modified Eagle’s medium (DMEM) supplemented with 10 % fetal bovine serum (Hyclone, Cytiva), penicillin (100 units/mL) and streptomycin (100 μg/mL) (Pen-Strep, Invitrogen) at 37°C in a 5 % CO 2 environment.

Techniques: Expressing, Incubation, Phospho-proteomics, Migration, Membrane

Journal: Free radical biology & medicine

Article Title: Redox regulation of cell migration via Nischarin S-glutathionylation

doi: 10.1016/j.freeradbiomed.2025.11.013

Figure Lengend Snippet: ( a ) A scheme of the G-PROV approach that enables functional analysis of glutathionylation on the protein of interest (POI): dehydroglutathione (dhG) is used to introduce a non-reducible glutathione modification on the purified engineered NISCH construct (enNISCH). Subsequently, the glutathione-modified NISCH construct is delivered to cells via fusogenic liposome, where the effect of glutathionylation on NISCH can be analyzed. ( b ) dhG modification on purified enNISCH. Purified enNISCH WT and C185S were incubated with dhG. The dhG-mediated glutathione modification was analyzed by glutathione antibody (n = 2). ( c ) Analysis of enNISCH delivered to MDA-MB-231 cells via fusogenic liposome. After incubating fusogenic liposomes containing His-enNISCH constructs (modified or non-modified by dhG) for 2 h, lysates were analyzed by Western blot (n = 2). ( d-e ) The migration and invasion assays. After incubating fusogenic liposomes for 2 h, MDA-MB-231 cells were analyzed by the wound-healing migration ( d , n = 6) and transwell invasion ( e , n = 2) assays. ( f ) The colony formation by enNISCH constructs. Fusogenic liposomes containing enNISCH WT or C185S were incubated with MDA-MB-231 cells on plates, and the number of colonies formed was counted after 15 days. Cells were incubated in DMEM containing 25 mM Glc ( d – e ) or 5 mM Glc ( f ). The statistical difference was analyzed by one-way ANOVA and Tukey’s post-hoc test ( b – e ) or two-way ANOVA and Sidak’s post-hoc test ( f ), where *p < 0.03, **p < 0.002, ***p < 0.0002, ****p < 0.0001.

Article Snippet: Human embryonic kidney 293 (HEK293) cell line stably expressing GS M4 (HEK293-GS M4) [ ], breast cancer cell line MDA-MB-231 (ATCC, HTB-26), and breast cancer cell line MCF-7 (ATCC, HTB-22) were maintained in Dulbecco’s modified Eagle’s medium (DMEM) supplemented with 10 % fetal bovine serum (Hyclone, Cytiva), penicillin (100 units/mL) and streptomycin (100 μg/mL) (Pen-Strep, Invitrogen) at 37°C in a 5 % CO 2 environment.

Techniques: Functional Assay, Introduce, Modification, Purification, Construct, Incubation, Liposomes, Western Blot, Migration

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