human breast cancer mcf7 (ATCC)

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ATCC human breast cancer mcf7

Effects of hypoxia on cell markers, morphology, and migration (A) Expression in breast cancer <t>MCF7</t> and MDA-MB-231 cells of E-cadherin (epithelial marker), fibronectin (epithelial marker), HIF-1α (hypoxia marker), β-catenin, and GLUT1. Cells were cultured at 37°C in 5% CO 2 atmosphere for normoxic treatment, and in 1% O 2 / 5% CO 2 / 94% N 2 atmosphere for hypoxic treatment. Cells were harvested, lysed in T-PER Reagent, and protein content was determined by BCA assay. Western blotting was performed as described in M&M. (B) Morphological changes under normoxic and hypoxic conditions. Cells (2 × 10 5 per well) were grown in 6-well plates for 24 h under the two conditions. Photos were taken by phase-contrast microscopy at 200× magnification. (C) Cell migration assessed by wound assay. Cell monolayers under the two conditions were scratched with pipette tip. Cells were washed with ice-cold 1× PBS and cultured in serum-free medium. Pictures of wounds were taken at 0 and 24 h by phase-contrast microscopy (100× magnification).

Human Breast Cancer Mcf7, supplied by ATCC, used in various techniques. Bioz Stars score: 99/100, based on 35160 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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1) Product Images from "Bisecting N-Acetylglucosamine Structures Inhibit Hypoxia-Induced Epithelial-Mesenchymal Transition in Breast Cancer Cells"

Article Title: Bisecting N-Acetylglucosamine Structures Inhibit Hypoxia-Induced Epithelial-Mesenchymal Transition in Breast Cancer Cells

Journal: Frontiers in Physiology

doi: 10.3389/fphys.2018.00210

Effects of hypoxia on cell markers, morphology, and migration (A) Expression in breast cancer MCF7 and MDA-MB-231 cells of E-cadherin (epithelial marker), fibronectin (epithelial marker), HIF-1α (hypoxia marker), β-catenin, and GLUT1. Cells were cultured at 37°C in 5% CO 2 atmosphere for normoxic treatment, and in 1% O 2 / 5% CO 2 / 94% N 2 atmosphere for hypoxic treatment. Cells were harvested, lysed in T-PER Reagent, and protein content was determined by BCA assay. Western blotting was performed as described in M&M. (B) Morphological changes under normoxic and hypoxic conditions. Cells (2 × 10 5 per well) were grown in 6-well plates for 24 h under the two conditions. Photos were taken by phase-contrast microscopy at 200× magnification. (C) Cell migration assessed by wound assay. Cell monolayers under the two conditions were scratched with pipette tip. Cells were washed with ice-cold 1× PBS and cultured in serum-free medium. Pictures of wounds were taken at 0 and 24 h by phase-contrast microscopy (100× magnification).

Figure Legend Snippet: Effects of hypoxia on cell markers, morphology, and migration (A) Expression in breast cancer MCF7 and MDA-MB-231 cells of E-cadherin (epithelial marker), fibronectin (epithelial marker), HIF-1α (hypoxia marker), β-catenin, and GLUT1. Cells were cultured at 37°C in 5% CO 2 atmosphere for normoxic treatment, and in 1% O 2 / 5% CO 2 / 94% N 2 atmosphere for hypoxic treatment. Cells were harvested, lysed in T-PER Reagent, and protein content was determined by BCA assay. Western blotting was performed as described in M&M. (B) Morphological changes under normoxic and hypoxic conditions. Cells (2 × 10 5 per well) were grown in 6-well plates for 24 h under the two conditions. Photos were taken by phase-contrast microscopy at 200× magnification. (C) Cell migration assessed by wound assay. Cell monolayers under the two conditions were scratched with pipette tip. Cells were washed with ice-cold 1× PBS and cultured in serum-free medium. Pictures of wounds were taken at 0 and 24 h by phase-contrast microscopy (100× magnification).

Techniques Used: Migration, Expressing, Marker, Cell Culture, BIA-KA, Western Blot, Microscopy, Transferring

MALDI-TOF-MS spectra of N-glycans from MCF7 cells MCF7 cells were cultured in 10-cm dishes under normoxic and hypoxic conditions, and N-glycans were separated and desalted as described in M&M. Lyophilized N-glycans were dissolved in MW, and an aliquot of mixture with DHB solution was spotted on MTP AnchorChip sample target and air-dried. MALTI-TOF-MS was performed in positive-ion mode. Experiments were performed in biological triplicate, and representative N-glycan spectra are shown. Peaks (signal-to-noise ratio > 5) were selected for relative proportion analysis. Detailed structures were analyzed using the GlycoWorkbench program. Proposed structures are indicated by m/z value.

Figure Legend Snippet: MALDI-TOF-MS spectra of N-glycans from MCF7 cells MCF7 cells were cultured in 10-cm dishes under normoxic and hypoxic conditions, and N-glycans were separated and desalted as described in M&M. Lyophilized N-glycans were dissolved in MW, and an aliquot of mixture with DHB solution was spotted on MTP AnchorChip sample target and air-dried. MALTI-TOF-MS was performed in positive-ion mode. Experiments were performed in biological triplicate, and representative N-glycan spectra are shown. Peaks (signal-to-noise ratio > 5) were selected for relative proportion analysis. Detailed structures were analyzed using the GlycoWorkbench program. Proposed structures are indicated by m/z value.

Techniques Used: Cell Culture, Glycoproteomics

Figure Legend Snippet: Relative proportions of various types of N-glycans in MCF7 and MDA-MB-231 cells under normoxia and hypoxia.

Techniques Used:

Variation of fine glycan structures detected by lectin microarray analysis (A) Variation of levels of glycans from MCF7 (upper) and MDA-MB-231 (lower) cells, detected by 37 lectins, is presented as a heatmap. Lectin microarray analysis was performed as described as M&M. Red: fluorescence signal activation. Green: signal inhibition. Black: missing data. (B) Altered glycan levels evaluated by lectin histochemistry. Four lectins (Con A, MAL-I, LCA, PHA-E) were applied, and lectin histochemistry was performed as described in M&M. Signals are shown from merge images of Cy3-conjugated lectins and DAPI staining of nuclei in MCF7 (left) and MDA-MB-231 (right) under normoxic and hypoxic conditions (60× magnification). (C) Expression in MCF7 and MDA-MB-231 cells of HIF-1α, MGAT3, and tubulin.

Figure Legend Snippet: Variation of fine glycan structures detected by lectin microarray analysis (A) Variation of levels of glycans from MCF7 (upper) and MDA-MB-231 (lower) cells, detected by 37 lectins, is presented as a heatmap. Lectin microarray analysis was performed as described as M&M. Red: fluorescence signal activation. Green: signal inhibition. Black: missing data. (B) Altered glycan levels evaluated by lectin histochemistry. Four lectins (Con A, MAL-I, LCA, PHA-E) were applied, and lectin histochemistry was performed as described in M&M. Signals are shown from merge images of Cy3-conjugated lectins and DAPI staining of nuclei in MCF7 (left) and MDA-MB-231 (right) under normoxic and hypoxic conditions (60× magnification). (C) Expression in MCF7 and MDA-MB-231 cells of HIF-1α, MGAT3, and tubulin.

Techniques Used: Glycoproteomics, Microarray, Fluorescence, Activation Assay, Inhibition, Staining, Expressing

Figure Legend Snippet: Differential glycopatterns in normoxia- vs. hypoxia-treated MCF7 cells revealed by lectin microarray analysis.

Techniques Used: Microarray

MGAT3 overexpression suppresses hypoxia-induced EMT in MCF7 cells (A) MGAT3 expression in mock- and MGAT3-transfected MCF7 cells. Cells were stably transduced with a GFP-marked lentivirus carrying mock gene or MGAT3 gene, harvested, and lysed in T-PER Reagent. Western blotting was performed as described in M&M using anti-MGAT3 and anti-GFP antibody. (B) Levels of bisecting GlcNAc structures in mock- and MGAT3-transfectants. Whole cell lysates of the two transfectants were subjected to PHA-E lectin blotting as described in M&M. (C) Proliferation of transfectant cells. The two transfectants were cultured for 24, 36, 48, 60, and 72 h, and proliferation was assessed by MTS assay. (E) Colony formation ability. The two transfectants (2500 cells each) were cultured in 6-cm dishes for 1–2 week, fixed, stained with crystal violet solution, and photographed. Acetic acid was added to dissolve crystal violet, and OD 595 was determined (D) . * p < 0.05; *** p < 0.001. (F) Cell migration. Migration assays of the two transfectants under normoxic and hypoxic conditions were performed as described in M&M, and relative migration rate was shown (H) . * p < 0.05. (G) Expression of HIF-1α, MGAT3, AKT, p-AKT, E-cadherin, fibronectin, and tubulin in the two transfectants under normoxic and hypoxic conditions. Cells were cultured as described in Figure , harvested, and lysed in T-PER Reagent. Protein content was determined by BCA assay. Western blotting was performed as described in M&M.

Figure Legend Snippet: MGAT3 overexpression suppresses hypoxia-induced EMT in MCF7 cells (A) MGAT3 expression in mock- and MGAT3-transfected MCF7 cells. Cells were stably transduced with a GFP-marked lentivirus carrying mock gene or MGAT3 gene, harvested, and lysed in T-PER Reagent. Western blotting was performed as described in M&M using anti-MGAT3 and anti-GFP antibody. (B) Levels of bisecting GlcNAc structures in mock- and MGAT3-transfectants. Whole cell lysates of the two transfectants were subjected to PHA-E lectin blotting as described in M&M. (C) Proliferation of transfectant cells. The two transfectants were cultured for 24, 36, 48, 60, and 72 h, and proliferation was assessed by MTS assay. (E) Colony formation ability. The two transfectants (2500 cells each) were cultured in 6-cm dishes for 1–2 week, fixed, stained with crystal violet solution, and photographed. Acetic acid was added to dissolve crystal violet, and OD 595 was determined (D) . * p < 0.05; *** p < 0.001. (F) Cell migration. Migration assays of the two transfectants under normoxic and hypoxic conditions were performed as described in M&M, and relative migration rate was shown (H) . * p < 0.05. (G) Expression of HIF-1α, MGAT3, AKT, p-AKT, E-cadherin, fibronectin, and tubulin in the two transfectants under normoxic and hypoxic conditions. Cells were cultured as described in Figure , harvested, and lysed in T-PER Reagent. Protein content was determined by BCA assay. Western blotting was performed as described in M&M.

Techniques Used: Over Expression, Expressing, Transfection, Stable Transfection, Transduction, Western Blot, Cell Culture, MTS Assay, Staining, Migration, BIA-KA

MGAT3 knockdown promotes hypoxia-induced EMT in MCF7 cells (A) MGAT3 expression in mock- and MGAT3-shRNA-transfected MCF7 cells. Cells were stably transduced with lentivirus carrying anti-MGAT3 shRNAs (MCF7/shMGAT3-1/2) or shNC (MCF7/mock), harvested, and lysed in T-PER Reagent. Western blotting was performed as described in M&M. (B) Cell proliferation. The two transfectants were cultured for 24, 36, 48, 60, and 72 h, and proliferation was assessed by MTS assay. (C) Cell migration. Migration assays of the two transfectants under normoxic and hypoxic conditions were performed as described in M&M, and relative migration rate was shown (E) . * p < 0.05; ** p < 0.01. (D) Expression of HIF-1α, MGAT3, AKT, p-AKT, E-cadherin, fibronectin, β-catenin, and tubulin in the two transfectants under normoxic and hypoxic conditions. Cells were cultured as described in Figure , harvested, and lysed in T-PER Reagent. Protein content was determined by BCA assay. Western blotting was performed as described in M&M.

Figure Legend Snippet: MGAT3 knockdown promotes hypoxia-induced EMT in MCF7 cells (A) MGAT3 expression in mock- and MGAT3-shRNA-transfected MCF7 cells. Cells were stably transduced with lentivirus carrying anti-MGAT3 shRNAs (MCF7/shMGAT3-1/2) or shNC (MCF7/mock), harvested, and lysed in T-PER Reagent. Western blotting was performed as described in M&M. (B) Cell proliferation. The two transfectants were cultured for 24, 36, 48, 60, and 72 h, and proliferation was assessed by MTS assay. (C) Cell migration. Migration assays of the two transfectants under normoxic and hypoxic conditions were performed as described in M&M, and relative migration rate was shown (E) . * p < 0.05; ** p < 0.01. (D) Expression of HIF-1α, MGAT3, AKT, p-AKT, E-cadherin, fibronectin, β-catenin, and tubulin in the two transfectants under normoxic and hypoxic conditions. Cells were cultured as described in Figure , harvested, and lysed in T-PER Reagent. Protein content was determined by BCA assay. Western blotting was performed as described in M&M.

Techniques Used: Knockdown, Expressing, shRNA, Transfection, Stable Transfection, Transduction, Western Blot, Cell Culture, MTS Assay, Migration, BIA-KA

Multiple Displacement Amplification:

Article Title: IL-1β/IL-6 inflammatory crosstalk drives DNA methylation and histone remodeling associated with cytokine induced senescence in luminal breast cancer
Article Snippet: .. MCF-7 (CVCL_0031), MDA-MB-231 (CVCL_0062) and MCF-10 A (CVCL_0598) cell lines were procured from ATCC and cultured according to supplier’s protocol. ..

Article Title: LINC01537 Regulates Malignant Phenotypes and Predicts Prognosis in Breast Cancer, and Its Bioinformatics Analysis
Article Snippet: .. The normal human mammary epithelial cell line MCF10A and breast cancer cell lines MDA‐MB‐453, MDA‐MB‐231, BT549, MCF‐7, and SK‐BR‐3 were obtained from the American Type Culture Collection (ATCC). .. MCF‐7 and MDA‐MB‐231 cells were grown in RPMI‐1640, while MCF10A, BT549, and MDA‐MB‐453 were maintained in DMEM (Invitrogen).

Cell Culture:

Article Title: Design and synthesis of hydrolyzable coumarin–naphthalimide dual probes for NIR fluorescence detection and selective photodynamic activity in cancer cells
Article Snippet: .. Human embryonic kidney cells HEK-293 (ATCC CRL-1573), used as a healthy model, and MCF-7 adenocarcinoma breast cancer cells (ATCC HTB-22) were cultured in Dulbecco's Modified Eagle's Medium (DMEM) supplemented with 10% fetal bovine serum (FBS), 100 U mL −1 penicillin, and 100 μg mL −1 streptomycin at 37 °C, 5% CO 2 , and 100% humidity. ..

Modification:

Flow Cytometry:

Article Title: Polymerase iota coordinates adaptation to replication stress in BRCA2-deficient mammary epithelial cells.
Article Snippet: REAGENT or RESOURCE SOURCE IDENTIFIER Antibodies Rabbit monoclonal anti-FANCD2 (Clone EPR2302) Abcam Cat# ab108928, RRID: AB_10862535 Mouse monoclonal anti-phospho-Histone H3 Ser10 (Clone 6G3) Cell Signaling Technology Cat# 9706, RRID: AB_331748 Mouse monoclonal anti-Cyclin A2 (Clone 1C6C11) Proteintech Cat# 66391-1-Ig, RRID: AB_2881767 Rabbit polyclonal anti-53BP1 Cell Signaling Technology Cat# 4937, RRID: AB_10694558 Rat monoclonal anti-RPA32/RPA2 (Clone 4E4) Cell Signaling Technology Cat# 2208, RRID: AB_2238543 Mouse monoclonal anti-γH2AX Ser139 (Clone JBW301) Sigma-Aldrich (EMD Millipore) Cat# 05–636, RRID: AB_309864 Mouse monoclonal anti-DNA Polymerase iota (POLI, Clone OTI1H9) Abcam Cat# ab157244 Mouse monoclonal BV421-conjugated antiCD24 (Clone ML5) BD Biosciences Cat# 562789, RRID: AB_2737796 Mouse monoclonal Alexa Fluor 647-conjugated ani-EpCAM (Clone VU1D9) Cell Signaling Technology Cat# 5447, RRID: AB_10693458 Rabbit monoclonal anti-BRCA2 (Clone D9S6V) Cell Signaling Technology Cat# 10741, RRID: AB_2797730 Rabbit monoclonal anti-RAD52 (Clone EPR3464(2)) Abcam Cat# ab124971, RRID: AB_10971685 Mouse monoclonal anti-β-Actin (Clone 8H10D10) Cell Signaling Technology Cat# 3700, RRID: AB_2242334 Rabbit monoclonal anti-Vinculin (Clone E1E9V) Cell Signaling Technology Cat# 13901, RRID: AB_2728768 Mouse monoclonal anti-BrdU (Clone B44) BD Biosciences Cat# 347580, RRID: AB_400326 Rat monoclonal anti-BrdU (Clone BU1/75 (ICR1)) Abcam Cat# ab6326, RRID: AB_305426 Biological samples Human breast tissues from prophylactic mastectomies of BRCA2 mutation carriers Tissue Procurement Core, Cleveland Clinic N/A Chemicals, peptides, and recombinant proteins Aphidicolin Sigma-Aldrich Cat# 5047440001 Colcemid Gibco Cat# 15210 Potassium chloride Sigma Cat# P9327 Giemsa Sigma-Aldrich Cat# 32884 Cytochalasin-B Sigma Cat# C2743 CIdU (5-Chloro-2′-deoxyuridine) Sigma-Aldrich Cat# C6891 IdU (5-Iodo-2′-deoxyuridine) Sigma-Aldrich Cat# I7125 Lipofectamine RNAiMAX ThermoFisher Scientific Cat# 13778075 DAPI Invitrogen Cat# P36962 DMEM/F12 media ThermoFisher Scientific Cat# 11330057 Horse Serum ThermoFisher Scientific Cat# 16050122 Insulin Sigma Cat# I-1882 EGF Peprotech Cat# AF-100–15 Hydrocortisone Sigma Cat# H6909 (Continued on next page) 12 iScience 29, 117830, November 20, 2026 .. REAGENT or RESOURCE SOURCE IDENTIFIER Cholera toxin Sigma Aldrich Cat# C8052 Mammary Epithelial Cell Basal Medium ATCC Cat# PCS-600–030, Mammary Epithelial Cell Growth Kit ATCC Cat# PCS-600–040 DMEM/F-12 GlutaMAX ThermoFisher Scientific Cat# 10565018 L-glutamine ThermoFisher Scientific Cat# 25030081 Gentamycin/amphotericin-B ThermoFisher Scientific Cat# R01510 Critical commercial assays Click-iT EdU Alexa Fluor 488 flow cytometry assay kit Invitrogen Cat# C10425 Click-iT EdU Alexa Fluor 488 Imaging kit Invitrogen Cat# C10637 FxCycle Violet Ready Flow Reagent Invitrogen Cat# R37166 Live-or-Dye 510/551 Biotium Cat # 32012A MycoAlert Mycoplasma Detection Kit Lonza Cat# LT07–218 PierceTM Rapid Gold BCA Protein Assay Kit Thermo Scientific, Cat# A53225 RIPA buffer Cell Signaling Technology Cat# 9803/9806 Protease and phosphatase inhibitors Cell Signaling Technology Cat# 5872 PMSF Cell Signaling Technology Cat# 8553 SDS-PAGE Precast Protein Gels Bio-Rad Cat# 4561084 Clarity Max Western ECL Substrate Bio-Rad Cat# 1705062 Experimental models: Cell lines Human mammary epithelial cell line: MCF10A Cleveland Clinic Research, cell and media production core facility Cat# ATCC, CRL-10317 Primary Human Mammary Epithelial Cells: HMECs American Type Culture Collection (ATCC) Cat# ATCC, PCS-600–010 Primary BRCA2-mutant Human Mammary Epithelial Cells: BRCA2mut/- HMECs Tissue Procurement Core, Cleveland Clinic Caputo et al.55 Human mammary cancer cell line: MCF7 Cleveland Clinic Research, cell and media production core facility Cat# ATCC, HTB-22 Human cervical cancer cell line: HeLa Cleveland Clinic Research, cell and media production core facility Cat# ATCC, CRM-CCL-2 Oligonucleotides siRNA sequences Dharmacon Horizon Discovery Supplementary file, this paper Software and algorithms R (version 4.5.3) R Software https://www.r-project.org/ Seurat (version 5.4.0) Hao, Y. et al., Dictionary learning for integrative, multimodal and scalable singlecell analysis. ..

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human breast cancer mcf7 (ATCC)

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1) Product Images from "Bisecting N-Acetylglucosamine Structures Inhibit Hypoxia-Induced Epithelial-Mesenchymal Transition in Breast Cancer Cells"

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