human breast cancer cell lines parental mda mb 231  (ATCC)


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    ATCC human breast cancer cell lines parental mda mb 231
    (A) Representative immunoblots of 3 independent studies of ITG proteins in Nsi and DAP5-silenced <t>MB-231</t> cells. Dox induction of shRNAs for 4 days followed by immunoblot analysis of equal protein amounts. (B) Representative immunoblots of 3 independent studies of EMT biomarker protein levels in Nsi and DAP5-silenced 4T1 and MB-231 cells carried out as in (A). (C) Representative immunoblots of 3 independent studies of eIF4E and eIF2α proteins in Nsi and DAP5-silenced MB-231 cells carried out as in (A). (D) Representative immunoblots of 3 independent studies to test requirement for eIF3d cap binding activity for DAP5-dependent mRNAs in MB-231 cells. Cells were small interfering RNA (siRNA) silenced for eIF3d for 1 day and transfected with vectors expressing WT, α5, or α11 cap binding mutants of eIF3d for 2 days, and equal protein amounts analyzed using immunoblot. (E) Matrigel Transwell invasion assays performed with 4T1 and MB-231 cells silenced for 24 h with Nsi or DAP5 Dox-inducible shRNAs. Mean number of invading cells/field with SEM from 3 replicates for each cell line. See also . (F) Cell migration wound healing assays performed with 4T1 and MB-231 cells silenced for 24 h with Nsi or DAP5 Dox-inducible shRNAs. Time 0 represents 100% wound separation of the cell layer. Mean of percentage of migrated cell surface areas (percentage closure) with SEM from 3 replicates for each cell line. See also . (G) Induction of apoptosis by cell detachment (anoikis) determined in 4T1 and MB-231 cells silenced for 24 and 48 h for Nsi, DAP5, or eIF4GI, comparing cells maintained on adherent or ultra- low-adherence plates. Percentage cell apoptosis determined by annexin V-fluorescein isothiocyanate (FITC) staining and flow cytometry, quantified using FlowJo software. Mean with SEM of 3 independent studies per cell line. (H) Representative immunoblot of 3 independent studies of DAP5, eIF4G1, eIF3d, and control β-actin protein levels in Nsi or DAP5-silenced 4T1 and MB-231 cells, silenced for 2 days with Dox-inducible shRNAs. Data are represented as mean ± SEM. n.s., not significant. *p < 0.05, **p < 0.01, ***p < 0.001, and ****p < 0.0001 by unpaired parametric two-tailed t test.
    Human Breast Cancer Cell Lines Parental Mda Mb 231, supplied by ATCC, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Images

    1) Product Images from "Breast cancer cell mesenchymal transition and metastasis directed by DAP5/eIF3d-mediated selective mRNA translation"

    Article Title: Breast cancer cell mesenchymal transition and metastasis directed by DAP5/eIF3d-mediated selective mRNA translation

    Journal: Cell reports

    doi: 10.1016/j.celrep.2023.112646

    (A) Representative immunoblots of 3 independent studies of ITG proteins in Nsi and DAP5-silenced MB-231 cells. Dox induction of shRNAs for 4 days followed by immunoblot analysis of equal protein amounts. (B) Representative immunoblots of 3 independent studies of EMT biomarker protein levels in Nsi and DAP5-silenced 4T1 and MB-231 cells carried out as in (A). (C) Representative immunoblots of 3 independent studies of eIF4E and eIF2α proteins in Nsi and DAP5-silenced MB-231 cells carried out as in (A). (D) Representative immunoblots of 3 independent studies to test requirement for eIF3d cap binding activity for DAP5-dependent mRNAs in MB-231 cells. Cells were small interfering RNA (siRNA) silenced for eIF3d for 1 day and transfected with vectors expressing WT, α5, or α11 cap binding mutants of eIF3d for 2 days, and equal protein amounts analyzed using immunoblot. (E) Matrigel Transwell invasion assays performed with 4T1 and MB-231 cells silenced for 24 h with Nsi or DAP5 Dox-inducible shRNAs. Mean number of invading cells/field with SEM from 3 replicates for each cell line. See also . (F) Cell migration wound healing assays performed with 4T1 and MB-231 cells silenced for 24 h with Nsi or DAP5 Dox-inducible shRNAs. Time 0 represents 100% wound separation of the cell layer. Mean of percentage of migrated cell surface areas (percentage closure) with SEM from 3 replicates for each cell line. See also . (G) Induction of apoptosis by cell detachment (anoikis) determined in 4T1 and MB-231 cells silenced for 24 and 48 h for Nsi, DAP5, or eIF4GI, comparing cells maintained on adherent or ultra- low-adherence plates. Percentage cell apoptosis determined by annexin V-fluorescein isothiocyanate (FITC) staining and flow cytometry, quantified using FlowJo software. Mean with SEM of 3 independent studies per cell line. (H) Representative immunoblot of 3 independent studies of DAP5, eIF4G1, eIF3d, and control β-actin protein levels in Nsi or DAP5-silenced 4T1 and MB-231 cells, silenced for 2 days with Dox-inducible shRNAs. Data are represented as mean ± SEM. n.s., not significant. *p < 0.05, **p < 0.01, ***p < 0.001, and ****p < 0.0001 by unpaired parametric two-tailed t test.
    Figure Legend Snippet: (A) Representative immunoblots of 3 independent studies of ITG proteins in Nsi and DAP5-silenced MB-231 cells. Dox induction of shRNAs for 4 days followed by immunoblot analysis of equal protein amounts. (B) Representative immunoblots of 3 independent studies of EMT biomarker protein levels in Nsi and DAP5-silenced 4T1 and MB-231 cells carried out as in (A). (C) Representative immunoblots of 3 independent studies of eIF4E and eIF2α proteins in Nsi and DAP5-silenced MB-231 cells carried out as in (A). (D) Representative immunoblots of 3 independent studies to test requirement for eIF3d cap binding activity for DAP5-dependent mRNAs in MB-231 cells. Cells were small interfering RNA (siRNA) silenced for eIF3d for 1 day and transfected with vectors expressing WT, α5, or α11 cap binding mutants of eIF3d for 2 days, and equal protein amounts analyzed using immunoblot. (E) Matrigel Transwell invasion assays performed with 4T1 and MB-231 cells silenced for 24 h with Nsi or DAP5 Dox-inducible shRNAs. Mean number of invading cells/field with SEM from 3 replicates for each cell line. See also . (F) Cell migration wound healing assays performed with 4T1 and MB-231 cells silenced for 24 h with Nsi or DAP5 Dox-inducible shRNAs. Time 0 represents 100% wound separation of the cell layer. Mean of percentage of migrated cell surface areas (percentage closure) with SEM from 3 replicates for each cell line. See also . (G) Induction of apoptosis by cell detachment (anoikis) determined in 4T1 and MB-231 cells silenced for 24 and 48 h for Nsi, DAP5, or eIF4GI, comparing cells maintained on adherent or ultra- low-adherence plates. Percentage cell apoptosis determined by annexin V-fluorescein isothiocyanate (FITC) staining and flow cytometry, quantified using FlowJo software. Mean with SEM of 3 independent studies per cell line. (H) Representative immunoblot of 3 independent studies of DAP5, eIF4G1, eIF3d, and control β-actin protein levels in Nsi or DAP5-silenced 4T1 and MB-231 cells, silenced for 2 days with Dox-inducible shRNAs. Data are represented as mean ± SEM. n.s., not significant. *p < 0.05, **p < 0.01, ***p < 0.001, and ****p < 0.0001 by unpaired parametric two-tailed t test.

    Techniques Used: Western Blot, Biomarker Assay, Binding Assay, Activity Assay, Small Interfering RNA, Transfection, Expressing, Migration, Staining, Flow Cytometry, Software, Two Tailed Test

    (A) NOD/SCID/γ mice orthotopically injected with 5 × 10 6 MB-231 cells stably transformed to express Firefly luciferase and red fluorescent protein (RFP), Dox-inducible Nsi or shDAP5 RNAs. Silencing initiated by Dox addition to drinking water 28 days after tumor cell implantation, tumors ~100–150 mm 3 in size. At 49 days, mice were sacrificed, tumors and lungs collected. (B) Growth of tumors was recorded every 7 days by quantitative caliper measurement. n = 8 or 9 mice per group repeated twice. Volumes of non-silenced control compared with DAP5-silenced tumors was not statistically significant. (C) Primary tumors weighed at excision 49 days. Non-silenced control compared with DAP5-silenced tumors not statistically significant. n = 7–9 mice/group. Representative of 2 trials. (D) Equal protein amounts of whole tumor lysates at 49 days post-excision, subjected to immunoblot analysis for DAP5 and control actin levels. n = 4 tumors (of 8 or 9 mice) per arm were evaluated. (E) Representative Firefly bioluminescent images from 8 or 9 mice per group (3 shown), silenced for Nsi or DAP5 as in (A) immediately prior to sacrifice at 49 days. Repeated 3 times. (F) Lungs excised, subjected to bioluminescent imaging for RFP. Representative images of 8 or 9 mice per group (4 shown) of mice silenced for Nsi or DAP5 as in (A). Repeated twice. (G) Quantification of metastatic burden in lungs of mice silenced for Nsi or DAP5 as in (A). Total RFP fluorescent flux quantified (photons [ps]/sec), representative of total lung tumor burden, at 49 days. n = 8 or 9 mice per group. Repeated twice. (H) Representative H&E-stained images of lungs of 8 or 9 mice per group harvested at 49 days. Arrows indicate dark staining tumor masses. (I) Quantification of number of lung metastases (mets) per field at 1003 magnification of H&E-stained lungs as shown in (H). A minimum of 5 fields per lung were quantified from 9 mice/group, mean plus SEM. (J) DAP5-silenced tumors have reduced levels of EMT proteins. Immunoblot of equal protein amounts from control Nsi and shDAP5-silenced tumors harvested at 49 days. Representative results of 6 tumors are shown. Data are mean plus SEM. Statistical significance was assessed by two-way ANOVA with Dunnett post-ANOVA test determination for analysis of repeated measures. n.s., not significant. ***p < 0.001. See also and .
    Figure Legend Snippet: (A) NOD/SCID/γ mice orthotopically injected with 5 × 10 6 MB-231 cells stably transformed to express Firefly luciferase and red fluorescent protein (RFP), Dox-inducible Nsi or shDAP5 RNAs. Silencing initiated by Dox addition to drinking water 28 days after tumor cell implantation, tumors ~100–150 mm 3 in size. At 49 days, mice were sacrificed, tumors and lungs collected. (B) Growth of tumors was recorded every 7 days by quantitative caliper measurement. n = 8 or 9 mice per group repeated twice. Volumes of non-silenced control compared with DAP5-silenced tumors was not statistically significant. (C) Primary tumors weighed at excision 49 days. Non-silenced control compared with DAP5-silenced tumors not statistically significant. n = 7–9 mice/group. Representative of 2 trials. (D) Equal protein amounts of whole tumor lysates at 49 days post-excision, subjected to immunoblot analysis for DAP5 and control actin levels. n = 4 tumors (of 8 or 9 mice) per arm were evaluated. (E) Representative Firefly bioluminescent images from 8 or 9 mice per group (3 shown), silenced for Nsi or DAP5 as in (A) immediately prior to sacrifice at 49 days. Repeated 3 times. (F) Lungs excised, subjected to bioluminescent imaging for RFP. Representative images of 8 or 9 mice per group (4 shown) of mice silenced for Nsi or DAP5 as in (A). Repeated twice. (G) Quantification of metastatic burden in lungs of mice silenced for Nsi or DAP5 as in (A). Total RFP fluorescent flux quantified (photons [ps]/sec), representative of total lung tumor burden, at 49 days. n = 8 or 9 mice per group. Repeated twice. (H) Representative H&E-stained images of lungs of 8 or 9 mice per group harvested at 49 days. Arrows indicate dark staining tumor masses. (I) Quantification of number of lung metastases (mets) per field at 1003 magnification of H&E-stained lungs as shown in (H). A minimum of 5 fields per lung were quantified from 9 mice/group, mean plus SEM. (J) DAP5-silenced tumors have reduced levels of EMT proteins. Immunoblot of equal protein amounts from control Nsi and shDAP5-silenced tumors harvested at 49 days. Representative results of 6 tumors are shown. Data are mean plus SEM. Statistical significance was assessed by two-way ANOVA with Dunnett post-ANOVA test determination for analysis of repeated measures. n.s., not significant. ***p < 0.001. See also and .

    Techniques Used: Injection, Stable Transfection, Transformation Assay, Luciferase, Western Blot, Imaging, Staining

    (A) MB-231 cells (1 × 10 7 ) expressing RFP and Dox-inducible control Nsi or shRNA to DAP5 implanted in mammary fat pad of NOD/SCID/γ mice, Dox induced at 28 days, when tumors 100–150 mm 3 in size, maintained for 20 days, tumors with stroma excised, embedded, sectioned for immunofluorescence microscopy for RFP (red) and DAPI (blue) stained nuclei. Representative images of RFP expressing MB-231 cells beyond the periphery of tumor, two different tumors analyzed, 3 fields each. Scale bar, 200 μm. Arrows indicate direction of tumor cell stromal migration away from tumor capsule. (B) Quantitation of images in (A). Quantitation determined from 2 independent tumors, 3 fields each (n = 6), using ImageJ software. (C) Tumors obtained as in (A) from non-silenced Nsi control and DAP5-silenced tumor specimens sectioned, IHC stained for CD31 to highlight tumor vasculature. Representative sections shown from 5 different tumors. Scale bar, 1,000 μm. (D) Quantitation of CD31 stained MB-231 cells from images in (C). Quantitation determined from 3 different tumors, 3 fields each. (E) Schema of animal study for survival determination with DAP5 tumor-specific silencing. Animals subcutaneously injected in flank with 1 × 10 5 4T1-Nsi or 4T1-shDAP5 cells. At 12 days, Dox added to the drinking water to induce eIF4G2 mRNA silencing. (F) Tumor growth recorded every 2–4 days by quantitative caliper measurement until 22 days as control mice began to die. n = 8–10 mice per group, repeated twice. (G) Survival of mice with 4T1-Nsi or 4T1-shDAP5 tumors silenced starting at 13 days. n = 15 mice/group. Animals either died or were sacrificed when terminally moribund. Statistical significance was determined using unpaired non-parametric log rank Mantel-Cox test. Data are mean with SEM. Statistical significance (B, D, and F) assessed using two-tailed Student’s t test.
    Figure Legend Snippet: (A) MB-231 cells (1 × 10 7 ) expressing RFP and Dox-inducible control Nsi or shRNA to DAP5 implanted in mammary fat pad of NOD/SCID/γ mice, Dox induced at 28 days, when tumors 100–150 mm 3 in size, maintained for 20 days, tumors with stroma excised, embedded, sectioned for immunofluorescence microscopy for RFP (red) and DAPI (blue) stained nuclei. Representative images of RFP expressing MB-231 cells beyond the periphery of tumor, two different tumors analyzed, 3 fields each. Scale bar, 200 μm. Arrows indicate direction of tumor cell stromal migration away from tumor capsule. (B) Quantitation of images in (A). Quantitation determined from 2 independent tumors, 3 fields each (n = 6), using ImageJ software. (C) Tumors obtained as in (A) from non-silenced Nsi control and DAP5-silenced tumor specimens sectioned, IHC stained for CD31 to highlight tumor vasculature. Representative sections shown from 5 different tumors. Scale bar, 1,000 μm. (D) Quantitation of CD31 stained MB-231 cells from images in (C). Quantitation determined from 3 different tumors, 3 fields each. (E) Schema of animal study for survival determination with DAP5 tumor-specific silencing. Animals subcutaneously injected in flank with 1 × 10 5 4T1-Nsi or 4T1-shDAP5 cells. At 12 days, Dox added to the drinking water to induce eIF4G2 mRNA silencing. (F) Tumor growth recorded every 2–4 days by quantitative caliper measurement until 22 days as control mice began to die. n = 8–10 mice per group, repeated twice. (G) Survival of mice with 4T1-Nsi or 4T1-shDAP5 tumors silenced starting at 13 days. n = 15 mice/group. Animals either died or were sacrificed when terminally moribund. Statistical significance was determined using unpaired non-parametric log rank Mantel-Cox test. Data are mean with SEM. Statistical significance (B, D, and F) assessed using two-tailed Student’s t test.

    Techniques Used: Expressing, shRNA, Immunofluorescence, Microscopy, Staining, Migration, Quantitation Assay, Software, Injection, Two Tailed Test

    (A) Representative immunoblot of select translation factor proteins in equal amounts of lysates from less transformed parental MB-231 cells, and two increasingly more transformed and more metastatic variants, LM0 and ML2 cells. (B) Representative immunoblot of 3 independent studies of parental MB-231 cells and parental MB-231 cells stably transfected with DAP5 cDNA (MB-231 OE cells). (C) Representative light field microscopic images of trypan blue stained parental MB-231 cells and MB-231 OE cells. n = 3 independent plating of cells, images representative from 12 different fields chosen at random. (D) Matrigel Transwell invasion assays performed with parental MB-231 cells or MB-231 OE cells, carried out as in . Results represent the mean of invading cells/field with SEM from 3 independent studies. Statistical analysis by unpaired two-tailed t test. (E) Cell migration wound healing assay. performed with parental MB-231 cells and MB-231 OE cells carried out as in . Statistical analysis by unpaired two-tailed t test. (F) Mice were injected in the retro-orbital (RO) sinus with 10 3 parental MB-231 cells or MB-231 OE cells, both expressing Firefly luciferase. Ten days later representative Firefly bioluminescent images were obtained from 3–5 mice per group (3 shown). Repeated 2 times. (G) Lungs excised at 10 days post-RO injection of mice described in (E), subjected to bioluminescent imaging for Firefly luciferase, repeated twice. (H) Quantification of metastatic burden in lungs of mice from (E). Total luciferase fluorescent flux quantified (photons [ps]/sec). *p < 0.05 by unpaired two-tailed t-test.
    Figure Legend Snippet: (A) Representative immunoblot of select translation factor proteins in equal amounts of lysates from less transformed parental MB-231 cells, and two increasingly more transformed and more metastatic variants, LM0 and ML2 cells. (B) Representative immunoblot of 3 independent studies of parental MB-231 cells and parental MB-231 cells stably transfected with DAP5 cDNA (MB-231 OE cells). (C) Representative light field microscopic images of trypan blue stained parental MB-231 cells and MB-231 OE cells. n = 3 independent plating of cells, images representative from 12 different fields chosen at random. (D) Matrigel Transwell invasion assays performed with parental MB-231 cells or MB-231 OE cells, carried out as in . Results represent the mean of invading cells/field with SEM from 3 independent studies. Statistical analysis by unpaired two-tailed t test. (E) Cell migration wound healing assay. performed with parental MB-231 cells and MB-231 OE cells carried out as in . Statistical analysis by unpaired two-tailed t test. (F) Mice were injected in the retro-orbital (RO) sinus with 10 3 parental MB-231 cells or MB-231 OE cells, both expressing Firefly luciferase. Ten days later representative Firefly bioluminescent images were obtained from 3–5 mice per group (3 shown). Repeated 2 times. (G) Lungs excised at 10 days post-RO injection of mice described in (E), subjected to bioluminescent imaging for Firefly luciferase, repeated twice. (H) Quantification of metastatic burden in lungs of mice from (E). Total luciferase fluorescent flux quantified (photons [ps]/sec). *p < 0.05 by unpaired two-tailed t-test.

    Techniques Used: Western Blot, Transformation Assay, Stable Transfection, Transfection, Staining, Two Tailed Test, Migration, Wound Healing Assay, Injection, Expressing, Luciferase, Imaging

    breast cancer cell line mda mb  (ATCC)


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    ATCC breast cancer cell line mda mb
    Selectivity of compound 1c on nine human cancer cell types.
    Breast Cancer Cell Line Mda Mb, supplied by ATCC, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/breast cancer cell line mda mb/product/ATCC
    Average 86 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    breast cancer cell line mda mb - by Bioz Stars, 2024-03
    86/100 stars

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    1) Product Images from "Design, synthesis, molecular modelling and biological evaluation of novel 6-amino-5-cyano-2-thiopyrimidine derivatives as potent anticancer agents against leukemia and apoptotic inducers"

    Article Title: Design, synthesis, molecular modelling and biological evaluation of novel 6-amino-5-cyano-2-thiopyrimidine derivatives as potent anticancer agents against leukemia and apoptotic inducers

    Journal: Journal of Enzyme Inhibition and Medicinal Chemistry

    doi: 10.1080/14756366.2024.2304625


    Figure Legend Snippet: Selectivity of compound 1c on nine human cancer cell types.

    Techniques Used:

    breast cancer cell line mda mb  (ATCC)


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    ATCC breast cancer cell line mda mb
    GFP-GOLPH3 localizes to mitochondrial fission sites after the recruitment of YFP-DRP1 in <t>MDA-MB-231</t> cells. MDA-MB-231 cells grown in 35 mm glass-bottom culture dishes were co-transfected to express GFP-GOLPH3 and YFP-DRP1. After 16 h, cells were incubated with MitoTracker TM Deep Red FM for 20 min at 37 °C, followed by the transfer of culture dishes to a microscope heating stage for time-lapse live-cell imaging. Images were acquired every ~0.86 s. ( A ) Representative imaged cell showing the cytoplasmic distribution of the labeled mitochondrial network (MitoTracker Deep Red; gray channel), GFP-GOLPH3 (green channel), and YFP-DRP1 (red channel). The fourth image depicts the superposition of the gray, green, and red channels (Merge). The boundary of the cell and nucleus (N) is marked with pale blue dashed lines. Bar, 10 μm. ( B ) Magnification of time-lapse images acquired at the indicated times of the cell region highlighted with an orange dashed line in the images shown in ( A ). The yellow arrows depict a mitochondrial fission event; the green arrows depict the recruitment and displacement of a GFP-GOLPH3 vesicle at the site of the mitochondrial fission highlighted by yellow arrows; and the red arrows depict the recruitment and displacement of YFP-DRP1 at the site of the mitochondrial fission highlighted by yellow arrows. Bar, 2 μm.
    Breast Cancer Cell Line Mda Mb, supplied by ATCC, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/breast cancer cell line mda mb/product/ATCC
    Average 86 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    breast cancer cell line mda mb - by Bioz Stars, 2024-03
    86/100 stars

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    1) Product Images from "GOLPH3 Participates in Mitochondrial Fission and Is Necessary to Sustain Bioenergetic Function in MDA-MB-231 Breast Cancer Cells"

    Article Title: GOLPH3 Participates in Mitochondrial Fission and Is Necessary to Sustain Bioenergetic Function in MDA-MB-231 Breast Cancer Cells

    Journal: Cells

    doi: 10.3390/cells13040316

    GFP-GOLPH3 localizes to mitochondrial fission sites after the recruitment of YFP-DRP1 in MDA-MB-231 cells. MDA-MB-231 cells grown in 35 mm glass-bottom culture dishes were co-transfected to express GFP-GOLPH3 and YFP-DRP1. After 16 h, cells were incubated with MitoTracker TM Deep Red FM for 20 min at 37 °C, followed by the transfer of culture dishes to a microscope heating stage for time-lapse live-cell imaging. Images were acquired every ~0.86 s. ( A ) Representative imaged cell showing the cytoplasmic distribution of the labeled mitochondrial network (MitoTracker Deep Red; gray channel), GFP-GOLPH3 (green channel), and YFP-DRP1 (red channel). The fourth image depicts the superposition of the gray, green, and red channels (Merge). The boundary of the cell and nucleus (N) is marked with pale blue dashed lines. Bar, 10 μm. ( B ) Magnification of time-lapse images acquired at the indicated times of the cell region highlighted with an orange dashed line in the images shown in ( A ). The yellow arrows depict a mitochondrial fission event; the green arrows depict the recruitment and displacement of a GFP-GOLPH3 vesicle at the site of the mitochondrial fission highlighted by yellow arrows; and the red arrows depict the recruitment and displacement of YFP-DRP1 at the site of the mitochondrial fission highlighted by yellow arrows. Bar, 2 μm.
    Figure Legend Snippet: GFP-GOLPH3 localizes to mitochondrial fission sites after the recruitment of YFP-DRP1 in MDA-MB-231 cells. MDA-MB-231 cells grown in 35 mm glass-bottom culture dishes were co-transfected to express GFP-GOLPH3 and YFP-DRP1. After 16 h, cells were incubated with MitoTracker TM Deep Red FM for 20 min at 37 °C, followed by the transfer of culture dishes to a microscope heating stage for time-lapse live-cell imaging. Images were acquired every ~0.86 s. ( A ) Representative imaged cell showing the cytoplasmic distribution of the labeled mitochondrial network (MitoTracker Deep Red; gray channel), GFP-GOLPH3 (green channel), and YFP-DRP1 (red channel). The fourth image depicts the superposition of the gray, green, and red channels (Merge). The boundary of the cell and nucleus (N) is marked with pale blue dashed lines. Bar, 10 μm. ( B ) Magnification of time-lapse images acquired at the indicated times of the cell region highlighted with an orange dashed line in the images shown in ( A ). The yellow arrows depict a mitochondrial fission event; the green arrows depict the recruitment and displacement of a GFP-GOLPH3 vesicle at the site of the mitochondrial fission highlighted by yellow arrows; and the red arrows depict the recruitment and displacement of YFP-DRP1 at the site of the mitochondrial fission highlighted by yellow arrows. Bar, 2 μm.

    Techniques Used: Transfection, Incubation, Microscopy, Live Cell Imaging, Labeling

    The depletion of GOLPH3 in MDA-MB-231 cells disrupts the mitochondrial network. ( A ) The indicated cells grown on glass coverslips were fixed, permeabilized, and decorated with mouse monoclonal antibodies against the mitochondrial protein TOM20. Stained cells were examined by fluorescence microscopy, and images were obtained in z-stack. The inset at the lower right corner of each image is a magnification of the respective region highlighted with a red dashed box. Bar, 10 μm. ( B , C ) Quantification of the number of mitochondrial profiles per cell area ( B ) and mitochondrial volume per cell area ( C ) performed using the 3D counter plugin of the ImageJ program. Bars represent the mean ± SEM (n = 3 independent experiments; n 1 = 15 cells, n 2 = 15 cells, n 3 = 26 cells). * p < 0.05; *** p < 0.001.
    Figure Legend Snippet: The depletion of GOLPH3 in MDA-MB-231 cells disrupts the mitochondrial network. ( A ) The indicated cells grown on glass coverslips were fixed, permeabilized, and decorated with mouse monoclonal antibodies against the mitochondrial protein TOM20. Stained cells were examined by fluorescence microscopy, and images were obtained in z-stack. The inset at the lower right corner of each image is a magnification of the respective region highlighted with a red dashed box. Bar, 10 μm. ( B , C ) Quantification of the number of mitochondrial profiles per cell area ( B ) and mitochondrial volume per cell area ( C ) performed using the 3D counter plugin of the ImageJ program. Bars represent the mean ± SEM (n = 3 independent experiments; n 1 = 15 cells, n 2 = 15 cells, n 3 = 26 cells). * p < 0.05; *** p < 0.001.

    Techniques Used: Staining, Fluorescence, Microscopy

    The depletion of GOLPH3 in MDA-MB-231 cells disrupts the outer mitochondrial membrane. The indicated cells were processed for transmission electron microscopy, and the images obtained ( A ) were analyzed to quantify outer mitochondrial membrane integrity ( B ) and electron density ( C ) using morphometric mitochondrial analyses implemented in Fiji software (version 2.1.0/1.53c). The orange arrow heads depict loss of the integrity of the mitochondrial outer membrane. Bar, 500 μm. Bars represent the mean ± SEM (n = 3 independent experiments; n 1 = 6 cells, n 2 = 6 cells, n 3 = 6 cells). * p < 0.05; ns: not statistically significant.
    Figure Legend Snippet: The depletion of GOLPH3 in MDA-MB-231 cells disrupts the outer mitochondrial membrane. The indicated cells were processed for transmission electron microscopy, and the images obtained ( A ) were analyzed to quantify outer mitochondrial membrane integrity ( B ) and electron density ( C ) using morphometric mitochondrial analyses implemented in Fiji software (version 2.1.0/1.53c). The orange arrow heads depict loss of the integrity of the mitochondrial outer membrane. Bar, 500 μm. Bars represent the mean ± SEM (n = 3 independent experiments; n 1 = 6 cells, n 2 = 6 cells, n 3 = 6 cells). * p < 0.05; ns: not statistically significant.

    Techniques Used: Membrane, Transmission Assay, Electron Microscopy, Software

    The depletion of GOLPH3 in MDA-MB-231 cells modifies the levels of mitochondrial fusion and fission proteins. ( A , C ) Detergent-soluble extracts were prepared from the indicated cells grown in 6-well plates, and the samples were processed by SDS-PAGE and immunoblot analysis using antibodies to detect the proteins indicated on the right involved in mitochondrial fusion ( A ) or mitochondrial fission ( C ). The immunoblot signal of anti-β-actin was used as the loading control. The position of molecular mass markers is indicated on the left. ( B , D ) Quantification of the respective immunoblot signal as shown in A and C. In C and D, pDRP1 denotes DRP1 phosphorylated at serine 616. Bars represent the mean ± SEM (n = 3). * p < 0.05; ** p < 0.01; ns: not statistically significant.
    Figure Legend Snippet: The depletion of GOLPH3 in MDA-MB-231 cells modifies the levels of mitochondrial fusion and fission proteins. ( A , C ) Detergent-soluble extracts were prepared from the indicated cells grown in 6-well plates, and the samples were processed by SDS-PAGE and immunoblot analysis using antibodies to detect the proteins indicated on the right involved in mitochondrial fusion ( A ) or mitochondrial fission ( C ). The immunoblot signal of anti-β-actin was used as the loading control. The position of molecular mass markers is indicated on the left. ( B , D ) Quantification of the respective immunoblot signal as shown in A and C. In C and D, pDRP1 denotes DRP1 phosphorylated at serine 616. Bars represent the mean ± SEM (n = 3). * p < 0.05; ** p < 0.01; ns: not statistically significant.

    Techniques Used: SDS Page, Western Blot

    GOLPH3 depletion in MDA-MB-231 cells results in decreased mitochondrial bioenergetics capacity and increased ROS content. ( A ) Protein extracts were prepared from the indicated cells, and ATP content was quantified using a luminescence assay (n = 15). ( B ) ATP production in a mitochondria-enriched fraction from the indicated cells incubated with oxidative substrates was quantified using a luminescence assay as in ( A ) (n = 5). ( C , D ) Mitochondrial membrane potential was quantified by incubating the indicated cells with the fluorescent dyes MitoTracker TM Red CMXRos (n = 3) (C) or with TMRE (n = 3) ( D ). ( E ) Oxygen consumption was quantified in the indicated cells using a fluorescent assay of cells incubated with an oxygen-quenchable fluorescent compound (n = 5). ( F ) ROS levels were quantified in the indicated cells using the fluorescent dye CM-H2DCFDA (n = 4). Bars represent the mean ± SEM. * p < 0.05; *** p < 0.001.
    Figure Legend Snippet: GOLPH3 depletion in MDA-MB-231 cells results in decreased mitochondrial bioenergetics capacity and increased ROS content. ( A ) Protein extracts were prepared from the indicated cells, and ATP content was quantified using a luminescence assay (n = 15). ( B ) ATP production in a mitochondria-enriched fraction from the indicated cells incubated with oxidative substrates was quantified using a luminescence assay as in ( A ) (n = 5). ( C , D ) Mitochondrial membrane potential was quantified by incubating the indicated cells with the fluorescent dyes MitoTracker TM Red CMXRos (n = 3) (C) or with TMRE (n = 3) ( D ). ( E ) Oxygen consumption was quantified in the indicated cells using a fluorescent assay of cells incubated with an oxygen-quenchable fluorescent compound (n = 5). ( F ) ROS levels were quantified in the indicated cells using the fluorescent dye CM-H2DCFDA (n = 4). Bars represent the mean ± SEM. * p < 0.05; *** p < 0.001.

    Techniques Used: Luminescence Assay, Incubation, Membrane, Fluorescence

    triple negative breast cancer mda mb 231 cell line  (ATCC)


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    ATCC triple negative breast cancer mda mb 231 cell line
    Effect of compounds on cancer cell viability at 100 µM concentration after 72 h of incubation against human triple-negative <t>breast</t> <t>cancer</t> <t>MDA-MB-231</t> and human pancreatic carcinoma Panc-1 cell lines by MTT assay, n = 3.
    Triple Negative Breast Cancer Mda Mb 231 Cell Line, supplied by ATCC, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    1) Product Images from "The Effect of 4-(Dimethylamino)phenyl-5-oxopyrrolidines on Breast and Pancreatic Cancer Cell Colony Formation, Migration, and Growth of Tumor Spheroids"

    Article Title: The Effect of 4-(Dimethylamino)phenyl-5-oxopyrrolidines on Breast and Pancreatic Cancer Cell Colony Formation, Migration, and Growth of Tumor Spheroids

    Journal: International Journal of Molecular Sciences

    doi: 10.3390/ijms25031834

    Effect of compounds on cancer cell viability at 100 µM concentration after 72 h of incubation against human triple-negative breast cancer MDA-MB-231 and human pancreatic carcinoma Panc-1 cell lines by MTT assay, n = 3.
    Figure Legend Snippet: Effect of compounds on cancer cell viability at 100 µM concentration after 72 h of incubation against human triple-negative breast cancer MDA-MB-231 and human pancreatic carcinoma Panc-1 cell lines by MTT assay, n = 3.

    Techniques Used: Concentration Assay, Incubation, MTT Assay

    EC 50 values of the most active compounds 3c , 3d , 5k , and 5l obtained by MTT assay ( A ) against triple-negative breast cancer MDA-MB231, pancreatic cancer Panc-1 cell lines, and human fibroblasts (HF) after 72 h of incubation. Dose–response curves for the most active compound 5k after 72 h of incubation with MDA-MB-231, Panc-1, and HF cells ( B ). Data points are experimental values (averages of three repeats), while the lines are the fit of the standard inhibition model with the Hill coefficient of 2.5. n = 3.
    Figure Legend Snippet: EC 50 values of the most active compounds 3c , 3d , 5k , and 5l obtained by MTT assay ( A ) against triple-negative breast cancer MDA-MB231, pancreatic cancer Panc-1 cell lines, and human fibroblasts (HF) after 72 h of incubation. Dose–response curves for the most active compound 5k after 72 h of incubation with MDA-MB-231, Panc-1, and HF cells ( B ). Data points are experimental values (averages of three repeats), while the lines are the fit of the standard inhibition model with the Hill coefficient of 2.5. n = 3.

    Techniques Used: MTT Assay, Incubation, Inhibition

    Effect of compounds 3c , 3d , 5k , and 5l on cell colony formation ( A ) and growth ( B ) of the human triple-negative breast cancer MDA-MB-231 cell line and cell colony formation ( C ) and growth ( D ) of the human pancreatic ductal carcinoma cell line, n = 3. Asterisks (*) indicate p < 0.05 compared with the control (untreated cells).
    Figure Legend Snippet: Effect of compounds 3c , 3d , 5k , and 5l on cell colony formation ( A ) and growth ( B ) of the human triple-negative breast cancer MDA-MB-231 cell line and cell colony formation ( C ) and growth ( D ) of the human pancreatic ductal carcinoma cell line, n = 3. Asterisks (*) indicate p < 0.05 compared with the control (untreated cells).

    Techniques Used:

    Effect of compounds 3c , 3d , 5k , and 5l on human triple-negative MDA-MB-231 and human pancreatic Panc-1 cell migration determined by ‘wound-healing’ assay. Photos of the ‘wound’ area (marked in yellow) in the MDA-MB-231 ( A ) and Panc-1 ( D ) monolayer at the beginning and the end of the experiment. The ‘wound’ area of MDA-MB-231 cells treated with 1 µM ( B ) and 2 µM ( C ) of the tested compounds at the end of the experiment, n = 3. The ‘wound’ area of Panc-1 cells treated with 1 µM ( E ) and 2 µM ( F ) of the tested compounds at the end of the experiment, n = 3. The scale bar indicates 100 µm. Asterisks (*) indicate p < 0.05 compared with the control (untreated cells).
    Figure Legend Snippet: Effect of compounds 3c , 3d , 5k , and 5l on human triple-negative MDA-MB-231 and human pancreatic Panc-1 cell migration determined by ‘wound-healing’ assay. Photos of the ‘wound’ area (marked in yellow) in the MDA-MB-231 ( A ) and Panc-1 ( D ) monolayer at the beginning and the end of the experiment. The ‘wound’ area of MDA-MB-231 cells treated with 1 µM ( B ) and 2 µM ( C ) of the tested compounds at the end of the experiment, n = 3. The ‘wound’ area of Panc-1 cells treated with 1 µM ( E ) and 2 µM ( F ) of the tested compounds at the end of the experiment, n = 3. The scale bar indicates 100 µm. Asterisks (*) indicate p < 0.05 compared with the control (untreated cells).

    Techniques Used: Migration, Wound Healing Assay

    Effect of compounds 3c , 3d , 5k , and 5l on 3D cell cultures. ( A ) Human pancreatic carcinoma Panc-1 spheroid size (brighter green color) and viability of cells in spheroids (lighter green color) at the end of the experiment. ( B ) Human triple-negative breast cancer MDA-MB-231 spheroid size (brighter blue color) and viability of cells in spheroids (lighter blue color) at the end of the experiment. ( C ) Photos of Panc-1 tumor spheroids at the beginning (Day 0), in the middle (Day 4), and at the end (Day 8) of the experiment (after incubation with 10 µM of the compounds). ( D ) Photos of MDA-MB-231 tumor spheroids at the beginning (Day 0), in the middle (Day 4), and at the end (Day 8) of the experiment (after incubation with 10 µM of the compounds). Asterisks (*) indicate p < 0.05 compared with the control (untreated spheroids); crosses (×) indicate means; inner dashes indicate medians; and whiskers indicate maximum and minimum values. Scale bars indicate 200 µm.
    Figure Legend Snippet: Effect of compounds 3c , 3d , 5k , and 5l on 3D cell cultures. ( A ) Human pancreatic carcinoma Panc-1 spheroid size (brighter green color) and viability of cells in spheroids (lighter green color) at the end of the experiment. ( B ) Human triple-negative breast cancer MDA-MB-231 spheroid size (brighter blue color) and viability of cells in spheroids (lighter blue color) at the end of the experiment. ( C ) Photos of Panc-1 tumor spheroids at the beginning (Day 0), in the middle (Day 4), and at the end (Day 8) of the experiment (after incubation with 10 µM of the compounds). ( D ) Photos of MDA-MB-231 tumor spheroids at the beginning (Day 0), in the middle (Day 4), and at the end (Day 8) of the experiment (after incubation with 10 µM of the compounds). Asterisks (*) indicate p < 0.05 compared with the control (untreated spheroids); crosses (×) indicate means; inner dashes indicate medians; and whiskers indicate maximum and minimum values. Scale bars indicate 200 µm.

    Techniques Used: Incubation

    human breast cancer cell lines mda mb 231  (ATCC)


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    ATCC human breast cancer cell lines mda mb 231
    Human Breast Cancer Cell Lines Mda Mb 231, supplied by ATCC, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    breast cancer epithelial cell lines mda mb 231  (ATCC)


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    ATCC breast cancer epithelial cell lines mda mb 231
    Breast Cancer Epithelial Cell Lines Mda Mb 231, supplied by ATCC, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    triple negative breast cancer cell line mda mb 231 cell line  (ATCC)


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    ATCC triple negative breast cancer cell line mda mb 231 cell line
    Cell uptake of [89Zr]Zr-DFO-NY003 and <t>[177Lu]Lu-DTPA-NY003</t> <t>in</t> <t>MDA-MB-231</t> ( A ) and ACHN ( B ) cells at different time points. (Block = co-incubation with non-radiolabeled NY003 and radiotracers for 48 and 72 h. Values were expressed as mean ± SD, n = 6. **Means P < 0.01, ns means not statistically significant)
    Triple Negative Breast Cancer Cell Line Mda Mb 231 Cell Line, supplied by ATCC, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    1) Product Images from "Preclinical evaluation of the theranostic potential of 89 Zr/ 177 Lu-labeled anti-TROP-2 antibody in triple-negative breast cancer model"

    Article Title: Preclinical evaluation of the theranostic potential of 89 Zr/ 177 Lu-labeled anti-TROP-2 antibody in triple-negative breast cancer model

    Journal: EJNMMI Radiopharmacy and Chemistry

    doi: 10.1186/s41181-023-00235-x

    Cell uptake of [89Zr]Zr-DFO-NY003 and [177Lu]Lu-DTPA-NY003 in MDA-MB-231 ( A ) and ACHN ( B ) cells at different time points. (Block = co-incubation with non-radiolabeled NY003 and radiotracers for 48 and 72 h. Values were expressed as mean ± SD, n = 6. **Means P < 0.01, ns means not statistically significant)
    Figure Legend Snippet: Cell uptake of [89Zr]Zr-DFO-NY003 and [177Lu]Lu-DTPA-NY003 in MDA-MB-231 ( A ) and ACHN ( B ) cells at different time points. (Block = co-incubation with non-radiolabeled NY003 and radiotracers for 48 and 72 h. Values were expressed as mean ± SD, n = 6. **Means P < 0.01, ns means not statistically significant)

    Techniques Used: Blocking Assay, Incubation

    The representative images of TROP-2 IHC staining of TROP-2 positive MDA-MB-231 ( A ) and TROP-2 negative ACHN ( B ) xenograft tumors, illustrating the weak to moderate specific TROP-2 over-expression of MDA-MB-231 tumor samples
    Figure Legend Snippet: The representative images of TROP-2 IHC staining of TROP-2 positive MDA-MB-231 ( A ) and TROP-2 negative ACHN ( B ) xenograft tumors, illustrating the weak to moderate specific TROP-2 over-expression of MDA-MB-231 tumor samples

    Techniques Used: Immunohistochemistry, Over Expression

    Micro-PET images in MDA-MB-231 tumor-bearing mice at different time points. A Representative MIP images of [ 89 Zr]Zr-DFO-NY003 (5.55–7.4 MBq) in MDA-MB-231 model at different time points p.i.. B Representative MIP images of [ 89 Zr]Zr-DFO-IgG (5.55 ~ 7.4 MBq) in the MDA-MB-231 model at different time points p.i.. C Representative MIP images of co-injection of [ 89 Zr]Zr-DFO-NY003 (5.55 ~ 7.4 MBq) and cold NY003 in MDA-MB-231 model at different time points p.i.. D–G Quantitative ROI analysis showed that the tumor uptake of [ 89 Zr]Zr-DFO-NY003 in MDA-MB-231 ( n = 3) was significantly higher than that of blocking and [ 89 Zr]Zr-DFO-IgG groups ( P < 0.001). The radioactive uptake of the heart (blood), liver, and kidney gradually decreased within the time tested
    Figure Legend Snippet: Micro-PET images in MDA-MB-231 tumor-bearing mice at different time points. A Representative MIP images of [ 89 Zr]Zr-DFO-NY003 (5.55–7.4 MBq) in MDA-MB-231 model at different time points p.i.. B Representative MIP images of [ 89 Zr]Zr-DFO-IgG (5.55 ~ 7.4 MBq) in the MDA-MB-231 model at different time points p.i.. C Representative MIP images of co-injection of [ 89 Zr]Zr-DFO-NY003 (5.55 ~ 7.4 MBq) and cold NY003 in MDA-MB-231 model at different time points p.i.. D–G Quantitative ROI analysis showed that the tumor uptake of [ 89 Zr]Zr-DFO-NY003 in MDA-MB-231 ( n = 3) was significantly higher than that of blocking and [ 89 Zr]Zr-DFO-IgG groups ( P < 0.001). The radioactive uptake of the heart (blood), liver, and kidney gradually decreased within the time tested

    Techniques Used: Micro-PET, Injection, Blocking Assay

    SPECT images in MDA-MB-231 and ACHN tumor-bearing mice at different time points p.i.. SPECT images showed specific high [ 177 Lu]Lu-DTPA-NY003 accumulation in tumors which can be blocked by cold NY003
    Figure Legend Snippet: SPECT images in MDA-MB-231 and ACHN tumor-bearing mice at different time points p.i.. SPECT images showed specific high [ 177 Lu]Lu-DTPA-NY003 accumulation in tumors which can be blocked by cold NY003

    Techniques Used: Single Photon Emission Computed Tomography

    triple negative breast cancer cell line mda mb 231 cell line  (ATCC)


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    ATCC triple negative breast cancer cell line mda mb 231 cell line
    Triple Negative Breast Cancer Cell Line Mda Mb 231 Cell Line, supplied by ATCC, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    mda mb 231 breast cancer cell lines  (ATCC)


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    ATCC mda mb 231 breast cancer cell lines
    LOX inhibition impairs osteoclast differentiation induced by <t>human</t> <t>MDA-MB</t> <t>231</t> and Hs 578T breast cancer cells. In vitro osteoclast differentiation of murine bone marrow cells treated with M-CSF + RANKL in combination with the conditioned medium from MDA-MB 231 or Hs 568T Ctrl, LOX(−) or LOXL2(−) cells. (A) TRAP staining of differentiated osteoclasts at day 7 of culture. Images shown are examples that best illustrate data obtained for each group. (B) Mature osteoclasts were quantified as multinucleated (more than 3 nuclei) TRAP-positive cells (n = 4). (C) Quantitative PCR determination of the relative levels of Rank , Trap , integrin β 3 ( itgb3 ) and cathepsin K ( cathK ) in murine bone marrow cells treated for 7 days with M-CSF + RANKL and the conditioned medium from MDA-MB 231 Ctrl, LOX(−), or LOXL2(−) cells (n = 9). Relative gene expression levels were normalized according to the C t value of the gene encoding the mouse ribosomal protein L32 *, **, ***: p < 0.05, 0.01 and 0.001, respectively.
    Mda Mb 231 Breast Cancer Cell Lines, supplied by ATCC, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    1) Product Images from "LOX, but not LOXL2, promotes bone metastasis formation and bone destruction in triple-negative breast cancer"

    Article Title: LOX, but not LOXL2, promotes bone metastasis formation and bone destruction in triple-negative breast cancer

    Journal: Journal of Bone Oncology

    doi: 10.1016/j.jbo.2024.100522

    LOX inhibition impairs osteoclast differentiation induced by human MDA-MB 231 and Hs 578T breast cancer cells. In vitro osteoclast differentiation of murine bone marrow cells treated with M-CSF + RANKL in combination with the conditioned medium from MDA-MB 231 or Hs 568T Ctrl, LOX(−) or LOXL2(−) cells. (A) TRAP staining of differentiated osteoclasts at day 7 of culture. Images shown are examples that best illustrate data obtained for each group. (B) Mature osteoclasts were quantified as multinucleated (more than 3 nuclei) TRAP-positive cells (n = 4). (C) Quantitative PCR determination of the relative levels of Rank , Trap , integrin β 3 ( itgb3 ) and cathepsin K ( cathK ) in murine bone marrow cells treated for 7 days with M-CSF + RANKL and the conditioned medium from MDA-MB 231 Ctrl, LOX(−), or LOXL2(−) cells (n = 9). Relative gene expression levels were normalized according to the C t value of the gene encoding the mouse ribosomal protein L32 *, **, ***: p < 0.05, 0.01 and 0.001, respectively.
    Figure Legend Snippet: LOX inhibition impairs osteoclast differentiation induced by human MDA-MB 231 and Hs 578T breast cancer cells. In vitro osteoclast differentiation of murine bone marrow cells treated with M-CSF + RANKL in combination with the conditioned medium from MDA-MB 231 or Hs 568T Ctrl, LOX(−) or LOXL2(−) cells. (A) TRAP staining of differentiated osteoclasts at day 7 of culture. Images shown are examples that best illustrate data obtained for each group. (B) Mature osteoclasts were quantified as multinucleated (more than 3 nuclei) TRAP-positive cells (n = 4). (C) Quantitative PCR determination of the relative levels of Rank , Trap , integrin β 3 ( itgb3 ) and cathepsin K ( cathK ) in murine bone marrow cells treated for 7 days with M-CSF + RANKL and the conditioned medium from MDA-MB 231 Ctrl, LOX(−), or LOXL2(−) cells (n = 9). Relative gene expression levels were normalized according to the C t value of the gene encoding the mouse ribosomal protein L32 *, **, ***: p < 0.05, 0.01 and 0.001, respectively.

    Techniques Used: Inhibition, In Vitro, Staining, Real-time Polymerase Chain Reaction, Expressing

    LOX overexpression in breast cancer cells increases the production of the pro-osteoclastic cytokine IL-6. (A) ELISA measurement of IL-6 in the conditioned medium from +/− βAPN-treated B02 LOX(+), LOX(−), LOXL2(+) or LOXL2(−) cells. Data are the mean ± SD of three independent experiments. (B) ELISA measurement of IL-6 in the conditioned medium from MDA-MB-231 Ctrl, LOX(−), or LOXL2(−) cells. Data are the mean ± SD of three independent experiments. (C) Correlation between IL-6 expression with that of LOX (left-hand panel) or LOXL2 (right-hand panel) in ER-negative primary mammary tumors (n = 78) from a cohort of breast cancer patients (GSE2034). Pearson correlation coefficients and P-values obtained with the PRISM 9 software are indicated. **, ***: p < 0.01 and 0.001, respectively.
    Figure Legend Snippet: LOX overexpression in breast cancer cells increases the production of the pro-osteoclastic cytokine IL-6. (A) ELISA measurement of IL-6 in the conditioned medium from +/− βAPN-treated B02 LOX(+), LOX(−), LOXL2(+) or LOXL2(−) cells. Data are the mean ± SD of three independent experiments. (B) ELISA measurement of IL-6 in the conditioned medium from MDA-MB-231 Ctrl, LOX(−), or LOXL2(−) cells. Data are the mean ± SD of three independent experiments. (C) Correlation between IL-6 expression with that of LOX (left-hand panel) or LOXL2 (right-hand panel) in ER-negative primary mammary tumors (n = 78) from a cohort of breast cancer patients (GSE2034). Pearson correlation coefficients and P-values obtained with the PRISM 9 software are indicated. **, ***: p < 0.01 and 0.001, respectively.

    Techniques Used: Over Expression, Enzyme-linked Immunosorbent Assay, Expressing, Software

    mda mb 468 breast cancer cell lines  (ATCC)


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    ATCC mda mb 468 breast cancer cell lines
    Mda Mb 468 Breast Cancer Cell Lines, supplied by ATCC, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    ATCC human breast cancer cell lines parental mda mb 231
    (A) Representative immunoblots of 3 independent studies of ITG proteins in Nsi and DAP5-silenced <t>MB-231</t> cells. Dox induction of shRNAs for 4 days followed by immunoblot analysis of equal protein amounts. (B) Representative immunoblots of 3 independent studies of EMT biomarker protein levels in Nsi and DAP5-silenced 4T1 and MB-231 cells carried out as in (A). (C) Representative immunoblots of 3 independent studies of eIF4E and eIF2α proteins in Nsi and DAP5-silenced MB-231 cells carried out as in (A). (D) Representative immunoblots of 3 independent studies to test requirement for eIF3d cap binding activity for DAP5-dependent mRNAs in MB-231 cells. Cells were small interfering RNA (siRNA) silenced for eIF3d for 1 day and transfected with vectors expressing WT, α5, or α11 cap binding mutants of eIF3d for 2 days, and equal protein amounts analyzed using immunoblot. (E) Matrigel Transwell invasion assays performed with 4T1 and MB-231 cells silenced for 24 h with Nsi or DAP5 Dox-inducible shRNAs. Mean number of invading cells/field with SEM from 3 replicates for each cell line. See also . (F) Cell migration wound healing assays performed with 4T1 and MB-231 cells silenced for 24 h with Nsi or DAP5 Dox-inducible shRNAs. Time 0 represents 100% wound separation of the cell layer. Mean of percentage of migrated cell surface areas (percentage closure) with SEM from 3 replicates for each cell line. See also . (G) Induction of apoptosis by cell detachment (anoikis) determined in 4T1 and MB-231 cells silenced for 24 and 48 h for Nsi, DAP5, or eIF4GI, comparing cells maintained on adherent or ultra- low-adherence plates. Percentage cell apoptosis determined by annexin V-fluorescein isothiocyanate (FITC) staining and flow cytometry, quantified using FlowJo software. Mean with SEM of 3 independent studies per cell line. (H) Representative immunoblot of 3 independent studies of DAP5, eIF4G1, eIF3d, and control β-actin protein levels in Nsi or DAP5-silenced 4T1 and MB-231 cells, silenced for 2 days with Dox-inducible shRNAs. Data are represented as mean ± SEM. n.s., not significant. *p < 0.05, **p < 0.01, ***p < 0.001, and ****p < 0.0001 by unpaired parametric two-tailed t test.
    Human Breast Cancer Cell Lines Parental Mda Mb 231, supplied by ATCC, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    ATCC breast cancer cell line mda mb
    Selectivity of compound 1c on nine human cancer cell types.
    Breast Cancer Cell Line Mda Mb, supplied by ATCC, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    ATCC triple negative breast cancer mda mb 231 cell line
    Effect of compounds on cancer cell viability at 100 µM concentration after 72 h of incubation against human triple-negative <t>breast</t> <t>cancer</t> <t>MDA-MB-231</t> and human pancreatic carcinoma Panc-1 cell lines by MTT assay, n = 3.
    Triple Negative Breast Cancer Mda Mb 231 Cell Line, supplied by ATCC, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    ATCC human breast cancer cell lines mda mb 231
    Effect of compounds on cancer cell viability at 100 µM concentration after 72 h of incubation against human triple-negative <t>breast</t> <t>cancer</t> <t>MDA-MB-231</t> and human pancreatic carcinoma Panc-1 cell lines by MTT assay, n = 3.
    Human Breast Cancer Cell Lines Mda Mb 231, supplied by ATCC, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    86
    ATCC breast cancer epithelial cell lines mda mb 231
    Effect of compounds on cancer cell viability at 100 µM concentration after 72 h of incubation against human triple-negative <t>breast</t> <t>cancer</t> <t>MDA-MB-231</t> and human pancreatic carcinoma Panc-1 cell lines by MTT assay, n = 3.
    Breast Cancer Epithelial Cell Lines Mda Mb 231, supplied by ATCC, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    ATCC triple negative breast cancer cell line mda mb 231 cell line
    Cell uptake of [89Zr]Zr-DFO-NY003 and <t>[177Lu]Lu-DTPA-NY003</t> <t>in</t> <t>MDA-MB-231</t> ( A ) and ACHN ( B ) cells at different time points. (Block = co-incubation with non-radiolabeled NY003 and radiotracers for 48 and 72 h. Values were expressed as mean ± SD, n = 6. **Means P < 0.01, ns means not statistically significant)
    Triple Negative Breast Cancer Cell Line Mda Mb 231 Cell Line, supplied by ATCC, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    ATCC mda mb 231 breast cancer cell lines
    LOX inhibition impairs osteoclast differentiation induced by <t>human</t> <t>MDA-MB</t> <t>231</t> and Hs 578T breast cancer cells. In vitro osteoclast differentiation of murine bone marrow cells treated with M-CSF + RANKL in combination with the conditioned medium from MDA-MB 231 or Hs 568T Ctrl, LOX(−) or LOXL2(−) cells. (A) TRAP staining of differentiated osteoclasts at day 7 of culture. Images shown are examples that best illustrate data obtained for each group. (B) Mature osteoclasts were quantified as multinucleated (more than 3 nuclei) TRAP-positive cells (n = 4). (C) Quantitative PCR determination of the relative levels of Rank , Trap , integrin β 3 ( itgb3 ) and cathepsin K ( cathK ) in murine bone marrow cells treated for 7 days with M-CSF + RANKL and the conditioned medium from MDA-MB 231 Ctrl, LOX(−), or LOXL2(−) cells (n = 9). Relative gene expression levels were normalized according to the C t value of the gene encoding the mouse ribosomal protein L32 *, **, ***: p < 0.05, 0.01 and 0.001, respectively.
    Mda Mb 231 Breast Cancer Cell Lines, supplied by ATCC, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    86
    ATCC mda mb 468 breast cancer cell lines
    LOX inhibition impairs osteoclast differentiation induced by <t>human</t> <t>MDA-MB</t> <t>231</t> and Hs 578T breast cancer cells. In vitro osteoclast differentiation of murine bone marrow cells treated with M-CSF + RANKL in combination with the conditioned medium from MDA-MB 231 or Hs 568T Ctrl, LOX(−) or LOXL2(−) cells. (A) TRAP staining of differentiated osteoclasts at day 7 of culture. Images shown are examples that best illustrate data obtained for each group. (B) Mature osteoclasts were quantified as multinucleated (more than 3 nuclei) TRAP-positive cells (n = 4). (C) Quantitative PCR determination of the relative levels of Rank , Trap , integrin β 3 ( itgb3 ) and cathepsin K ( cathK ) in murine bone marrow cells treated for 7 days with M-CSF + RANKL and the conditioned medium from MDA-MB 231 Ctrl, LOX(−), or LOXL2(−) cells (n = 9). Relative gene expression levels were normalized according to the C t value of the gene encoding the mouse ribosomal protein L32 *, **, ***: p < 0.05, 0.01 and 0.001, respectively.
    Mda Mb 468 Breast Cancer Cell Lines, supplied by ATCC, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Image Search Results


    (A) Representative immunoblots of 3 independent studies of ITG proteins in Nsi and DAP5-silenced MB-231 cells. Dox induction of shRNAs for 4 days followed by immunoblot analysis of equal protein amounts. (B) Representative immunoblots of 3 independent studies of EMT biomarker protein levels in Nsi and DAP5-silenced 4T1 and MB-231 cells carried out as in (A). (C) Representative immunoblots of 3 independent studies of eIF4E and eIF2α proteins in Nsi and DAP5-silenced MB-231 cells carried out as in (A). (D) Representative immunoblots of 3 independent studies to test requirement for eIF3d cap binding activity for DAP5-dependent mRNAs in MB-231 cells. Cells were small interfering RNA (siRNA) silenced for eIF3d for 1 day and transfected with vectors expressing WT, α5, or α11 cap binding mutants of eIF3d for 2 days, and equal protein amounts analyzed using immunoblot. (E) Matrigel Transwell invasion assays performed with 4T1 and MB-231 cells silenced for 24 h with Nsi or DAP5 Dox-inducible shRNAs. Mean number of invading cells/field with SEM from 3 replicates for each cell line. See also . (F) Cell migration wound healing assays performed with 4T1 and MB-231 cells silenced for 24 h with Nsi or DAP5 Dox-inducible shRNAs. Time 0 represents 100% wound separation of the cell layer. Mean of percentage of migrated cell surface areas (percentage closure) with SEM from 3 replicates for each cell line. See also . (G) Induction of apoptosis by cell detachment (anoikis) determined in 4T1 and MB-231 cells silenced for 24 and 48 h for Nsi, DAP5, or eIF4GI, comparing cells maintained on adherent or ultra- low-adherence plates. Percentage cell apoptosis determined by annexin V-fluorescein isothiocyanate (FITC) staining and flow cytometry, quantified using FlowJo software. Mean with SEM of 3 independent studies per cell line. (H) Representative immunoblot of 3 independent studies of DAP5, eIF4G1, eIF3d, and control β-actin protein levels in Nsi or DAP5-silenced 4T1 and MB-231 cells, silenced for 2 days with Dox-inducible shRNAs. Data are represented as mean ± SEM. n.s., not significant. *p < 0.05, **p < 0.01, ***p < 0.001, and ****p < 0.0001 by unpaired parametric two-tailed t test.

    Journal: Cell reports

    Article Title: Breast cancer cell mesenchymal transition and metastasis directed by DAP5/eIF3d-mediated selective mRNA translation

    doi: 10.1016/j.celrep.2023.112646

    Figure Lengend Snippet: (A) Representative immunoblots of 3 independent studies of ITG proteins in Nsi and DAP5-silenced MB-231 cells. Dox induction of shRNAs for 4 days followed by immunoblot analysis of equal protein amounts. (B) Representative immunoblots of 3 independent studies of EMT biomarker protein levels in Nsi and DAP5-silenced 4T1 and MB-231 cells carried out as in (A). (C) Representative immunoblots of 3 independent studies of eIF4E and eIF2α proteins in Nsi and DAP5-silenced MB-231 cells carried out as in (A). (D) Representative immunoblots of 3 independent studies to test requirement for eIF3d cap binding activity for DAP5-dependent mRNAs in MB-231 cells. Cells were small interfering RNA (siRNA) silenced for eIF3d for 1 day and transfected with vectors expressing WT, α5, or α11 cap binding mutants of eIF3d for 2 days, and equal protein amounts analyzed using immunoblot. (E) Matrigel Transwell invasion assays performed with 4T1 and MB-231 cells silenced for 24 h with Nsi or DAP5 Dox-inducible shRNAs. Mean number of invading cells/field with SEM from 3 replicates for each cell line. See also . (F) Cell migration wound healing assays performed with 4T1 and MB-231 cells silenced for 24 h with Nsi or DAP5 Dox-inducible shRNAs. Time 0 represents 100% wound separation of the cell layer. Mean of percentage of migrated cell surface areas (percentage closure) with SEM from 3 replicates for each cell line. See also . (G) Induction of apoptosis by cell detachment (anoikis) determined in 4T1 and MB-231 cells silenced for 24 and 48 h for Nsi, DAP5, or eIF4GI, comparing cells maintained on adherent or ultra- low-adherence plates. Percentage cell apoptosis determined by annexin V-fluorescein isothiocyanate (FITC) staining and flow cytometry, quantified using FlowJo software. Mean with SEM of 3 independent studies per cell line. (H) Representative immunoblot of 3 independent studies of DAP5, eIF4G1, eIF3d, and control β-actin protein levels in Nsi or DAP5-silenced 4T1 and MB-231 cells, silenced for 2 days with Dox-inducible shRNAs. Data are represented as mean ± SEM. n.s., not significant. *p < 0.05, **p < 0.01, ***p < 0.001, and ****p < 0.0001 by unpaired parametric two-tailed t test.

    Article Snippet: The human breast cancer cell lines parental MDA-MB-231, MDA-MB-231 LM0 and MDA-MB-231 LM2, and the murine mammary carcinoma cell line 4T1 were obtained from the American Type Culture Collection (ATCC, Manassas, VA, USA).

    Techniques: Western Blot, Biomarker Assay, Binding Assay, Activity Assay, Small Interfering RNA, Transfection, Expressing, Migration, Staining, Flow Cytometry, Software, Two Tailed Test

    (A) NOD/SCID/γ mice orthotopically injected with 5 × 10 6 MB-231 cells stably transformed to express Firefly luciferase and red fluorescent protein (RFP), Dox-inducible Nsi or shDAP5 RNAs. Silencing initiated by Dox addition to drinking water 28 days after tumor cell implantation, tumors ~100–150 mm 3 in size. At 49 days, mice were sacrificed, tumors and lungs collected. (B) Growth of tumors was recorded every 7 days by quantitative caliper measurement. n = 8 or 9 mice per group repeated twice. Volumes of non-silenced control compared with DAP5-silenced tumors was not statistically significant. (C) Primary tumors weighed at excision 49 days. Non-silenced control compared with DAP5-silenced tumors not statistically significant. n = 7–9 mice/group. Representative of 2 trials. (D) Equal protein amounts of whole tumor lysates at 49 days post-excision, subjected to immunoblot analysis for DAP5 and control actin levels. n = 4 tumors (of 8 or 9 mice) per arm were evaluated. (E) Representative Firefly bioluminescent images from 8 or 9 mice per group (3 shown), silenced for Nsi or DAP5 as in (A) immediately prior to sacrifice at 49 days. Repeated 3 times. (F) Lungs excised, subjected to bioluminescent imaging for RFP. Representative images of 8 or 9 mice per group (4 shown) of mice silenced for Nsi or DAP5 as in (A). Repeated twice. (G) Quantification of metastatic burden in lungs of mice silenced for Nsi or DAP5 as in (A). Total RFP fluorescent flux quantified (photons [ps]/sec), representative of total lung tumor burden, at 49 days. n = 8 or 9 mice per group. Repeated twice. (H) Representative H&E-stained images of lungs of 8 or 9 mice per group harvested at 49 days. Arrows indicate dark staining tumor masses. (I) Quantification of number of lung metastases (mets) per field at 1003 magnification of H&E-stained lungs as shown in (H). A minimum of 5 fields per lung were quantified from 9 mice/group, mean plus SEM. (J) DAP5-silenced tumors have reduced levels of EMT proteins. Immunoblot of equal protein amounts from control Nsi and shDAP5-silenced tumors harvested at 49 days. Representative results of 6 tumors are shown. Data are mean plus SEM. Statistical significance was assessed by two-way ANOVA with Dunnett post-ANOVA test determination for analysis of repeated measures. n.s., not significant. ***p < 0.001. See also and .

    Journal: Cell reports

    Article Title: Breast cancer cell mesenchymal transition and metastasis directed by DAP5/eIF3d-mediated selective mRNA translation

    doi: 10.1016/j.celrep.2023.112646

    Figure Lengend Snippet: (A) NOD/SCID/γ mice orthotopically injected with 5 × 10 6 MB-231 cells stably transformed to express Firefly luciferase and red fluorescent protein (RFP), Dox-inducible Nsi or shDAP5 RNAs. Silencing initiated by Dox addition to drinking water 28 days after tumor cell implantation, tumors ~100–150 mm 3 in size. At 49 days, mice were sacrificed, tumors and lungs collected. (B) Growth of tumors was recorded every 7 days by quantitative caliper measurement. n = 8 or 9 mice per group repeated twice. Volumes of non-silenced control compared with DAP5-silenced tumors was not statistically significant. (C) Primary tumors weighed at excision 49 days. Non-silenced control compared with DAP5-silenced tumors not statistically significant. n = 7–9 mice/group. Representative of 2 trials. (D) Equal protein amounts of whole tumor lysates at 49 days post-excision, subjected to immunoblot analysis for DAP5 and control actin levels. n = 4 tumors (of 8 or 9 mice) per arm were evaluated. (E) Representative Firefly bioluminescent images from 8 or 9 mice per group (3 shown), silenced for Nsi or DAP5 as in (A) immediately prior to sacrifice at 49 days. Repeated 3 times. (F) Lungs excised, subjected to bioluminescent imaging for RFP. Representative images of 8 or 9 mice per group (4 shown) of mice silenced for Nsi or DAP5 as in (A). Repeated twice. (G) Quantification of metastatic burden in lungs of mice silenced for Nsi or DAP5 as in (A). Total RFP fluorescent flux quantified (photons [ps]/sec), representative of total lung tumor burden, at 49 days. n = 8 or 9 mice per group. Repeated twice. (H) Representative H&E-stained images of lungs of 8 or 9 mice per group harvested at 49 days. Arrows indicate dark staining tumor masses. (I) Quantification of number of lung metastases (mets) per field at 1003 magnification of H&E-stained lungs as shown in (H). A minimum of 5 fields per lung were quantified from 9 mice/group, mean plus SEM. (J) DAP5-silenced tumors have reduced levels of EMT proteins. Immunoblot of equal protein amounts from control Nsi and shDAP5-silenced tumors harvested at 49 days. Representative results of 6 tumors are shown. Data are mean plus SEM. Statistical significance was assessed by two-way ANOVA with Dunnett post-ANOVA test determination for analysis of repeated measures. n.s., not significant. ***p < 0.001. See also and .

    Article Snippet: The human breast cancer cell lines parental MDA-MB-231, MDA-MB-231 LM0 and MDA-MB-231 LM2, and the murine mammary carcinoma cell line 4T1 were obtained from the American Type Culture Collection (ATCC, Manassas, VA, USA).

    Techniques: Injection, Stable Transfection, Transformation Assay, Luciferase, Western Blot, Imaging, Staining

    (A) MB-231 cells (1 × 10 7 ) expressing RFP and Dox-inducible control Nsi or shRNA to DAP5 implanted in mammary fat pad of NOD/SCID/γ mice, Dox induced at 28 days, when tumors 100–150 mm 3 in size, maintained for 20 days, tumors with stroma excised, embedded, sectioned for immunofluorescence microscopy for RFP (red) and DAPI (blue) stained nuclei. Representative images of RFP expressing MB-231 cells beyond the periphery of tumor, two different tumors analyzed, 3 fields each. Scale bar, 200 μm. Arrows indicate direction of tumor cell stromal migration away from tumor capsule. (B) Quantitation of images in (A). Quantitation determined from 2 independent tumors, 3 fields each (n = 6), using ImageJ software. (C) Tumors obtained as in (A) from non-silenced Nsi control and DAP5-silenced tumor specimens sectioned, IHC stained for CD31 to highlight tumor vasculature. Representative sections shown from 5 different tumors. Scale bar, 1,000 μm. (D) Quantitation of CD31 stained MB-231 cells from images in (C). Quantitation determined from 3 different tumors, 3 fields each. (E) Schema of animal study for survival determination with DAP5 tumor-specific silencing. Animals subcutaneously injected in flank with 1 × 10 5 4T1-Nsi or 4T1-shDAP5 cells. At 12 days, Dox added to the drinking water to induce eIF4G2 mRNA silencing. (F) Tumor growth recorded every 2–4 days by quantitative caliper measurement until 22 days as control mice began to die. n = 8–10 mice per group, repeated twice. (G) Survival of mice with 4T1-Nsi or 4T1-shDAP5 tumors silenced starting at 13 days. n = 15 mice/group. Animals either died or were sacrificed when terminally moribund. Statistical significance was determined using unpaired non-parametric log rank Mantel-Cox test. Data are mean with SEM. Statistical significance (B, D, and F) assessed using two-tailed Student’s t test.

    Journal: Cell reports

    Article Title: Breast cancer cell mesenchymal transition and metastasis directed by DAP5/eIF3d-mediated selective mRNA translation

    doi: 10.1016/j.celrep.2023.112646

    Figure Lengend Snippet: (A) MB-231 cells (1 × 10 7 ) expressing RFP and Dox-inducible control Nsi or shRNA to DAP5 implanted in mammary fat pad of NOD/SCID/γ mice, Dox induced at 28 days, when tumors 100–150 mm 3 in size, maintained for 20 days, tumors with stroma excised, embedded, sectioned for immunofluorescence microscopy for RFP (red) and DAPI (blue) stained nuclei. Representative images of RFP expressing MB-231 cells beyond the periphery of tumor, two different tumors analyzed, 3 fields each. Scale bar, 200 μm. Arrows indicate direction of tumor cell stromal migration away from tumor capsule. (B) Quantitation of images in (A). Quantitation determined from 2 independent tumors, 3 fields each (n = 6), using ImageJ software. (C) Tumors obtained as in (A) from non-silenced Nsi control and DAP5-silenced tumor specimens sectioned, IHC stained for CD31 to highlight tumor vasculature. Representative sections shown from 5 different tumors. Scale bar, 1,000 μm. (D) Quantitation of CD31 stained MB-231 cells from images in (C). Quantitation determined from 3 different tumors, 3 fields each. (E) Schema of animal study for survival determination with DAP5 tumor-specific silencing. Animals subcutaneously injected in flank with 1 × 10 5 4T1-Nsi or 4T1-shDAP5 cells. At 12 days, Dox added to the drinking water to induce eIF4G2 mRNA silencing. (F) Tumor growth recorded every 2–4 days by quantitative caliper measurement until 22 days as control mice began to die. n = 8–10 mice per group, repeated twice. (G) Survival of mice with 4T1-Nsi or 4T1-shDAP5 tumors silenced starting at 13 days. n = 15 mice/group. Animals either died or were sacrificed when terminally moribund. Statistical significance was determined using unpaired non-parametric log rank Mantel-Cox test. Data are mean with SEM. Statistical significance (B, D, and F) assessed using two-tailed Student’s t test.

    Article Snippet: The human breast cancer cell lines parental MDA-MB-231, MDA-MB-231 LM0 and MDA-MB-231 LM2, and the murine mammary carcinoma cell line 4T1 were obtained from the American Type Culture Collection (ATCC, Manassas, VA, USA).

    Techniques: Expressing, shRNA, Immunofluorescence, Microscopy, Staining, Migration, Quantitation Assay, Software, Injection, Two Tailed Test

    (A) Representative immunoblot of select translation factor proteins in equal amounts of lysates from less transformed parental MB-231 cells, and two increasingly more transformed and more metastatic variants, LM0 and ML2 cells. (B) Representative immunoblot of 3 independent studies of parental MB-231 cells and parental MB-231 cells stably transfected with DAP5 cDNA (MB-231 OE cells). (C) Representative light field microscopic images of trypan blue stained parental MB-231 cells and MB-231 OE cells. n = 3 independent plating of cells, images representative from 12 different fields chosen at random. (D) Matrigel Transwell invasion assays performed with parental MB-231 cells or MB-231 OE cells, carried out as in . Results represent the mean of invading cells/field with SEM from 3 independent studies. Statistical analysis by unpaired two-tailed t test. (E) Cell migration wound healing assay. performed with parental MB-231 cells and MB-231 OE cells carried out as in . Statistical analysis by unpaired two-tailed t test. (F) Mice were injected in the retro-orbital (RO) sinus with 10 3 parental MB-231 cells or MB-231 OE cells, both expressing Firefly luciferase. Ten days later representative Firefly bioluminescent images were obtained from 3–5 mice per group (3 shown). Repeated 2 times. (G) Lungs excised at 10 days post-RO injection of mice described in (E), subjected to bioluminescent imaging for Firefly luciferase, repeated twice. (H) Quantification of metastatic burden in lungs of mice from (E). Total luciferase fluorescent flux quantified (photons [ps]/sec). *p < 0.05 by unpaired two-tailed t-test.

    Journal: Cell reports

    Article Title: Breast cancer cell mesenchymal transition and metastasis directed by DAP5/eIF3d-mediated selective mRNA translation

    doi: 10.1016/j.celrep.2023.112646

    Figure Lengend Snippet: (A) Representative immunoblot of select translation factor proteins in equal amounts of lysates from less transformed parental MB-231 cells, and two increasingly more transformed and more metastatic variants, LM0 and ML2 cells. (B) Representative immunoblot of 3 independent studies of parental MB-231 cells and parental MB-231 cells stably transfected with DAP5 cDNA (MB-231 OE cells). (C) Representative light field microscopic images of trypan blue stained parental MB-231 cells and MB-231 OE cells. n = 3 independent plating of cells, images representative from 12 different fields chosen at random. (D) Matrigel Transwell invasion assays performed with parental MB-231 cells or MB-231 OE cells, carried out as in . Results represent the mean of invading cells/field with SEM from 3 independent studies. Statistical analysis by unpaired two-tailed t test. (E) Cell migration wound healing assay. performed with parental MB-231 cells and MB-231 OE cells carried out as in . Statistical analysis by unpaired two-tailed t test. (F) Mice were injected in the retro-orbital (RO) sinus with 10 3 parental MB-231 cells or MB-231 OE cells, both expressing Firefly luciferase. Ten days later representative Firefly bioluminescent images were obtained from 3–5 mice per group (3 shown). Repeated 2 times. (G) Lungs excised at 10 days post-RO injection of mice described in (E), subjected to bioluminescent imaging for Firefly luciferase, repeated twice. (H) Quantification of metastatic burden in lungs of mice from (E). Total luciferase fluorescent flux quantified (photons [ps]/sec). *p < 0.05 by unpaired two-tailed t-test.

    Article Snippet: The human breast cancer cell lines parental MDA-MB-231, MDA-MB-231 LM0 and MDA-MB-231 LM2, and the murine mammary carcinoma cell line 4T1 were obtained from the American Type Culture Collection (ATCC, Manassas, VA, USA).

    Techniques: Western Blot, Transformation Assay, Stable Transfection, Transfection, Staining, Two Tailed Test, Migration, Wound Healing Assay, Injection, Expressing, Luciferase, Imaging

    Journal: Journal of Enzyme Inhibition and Medicinal Chemistry

    Article Title: Design, synthesis, molecular modelling and biological evaluation of novel 6-amino-5-cyano-2-thiopyrimidine derivatives as potent anticancer agents against leukemia and apoptotic inducers

    doi: 10.1080/14756366.2024.2304625

    Figure Lengend Snippet: Selectivity of compound 1c on nine human cancer cell types.

    Article Snippet: Compound 4k showed significant anticancer activity against Non-Small Cell Lung Cancer cell line HOP-92 with growth percentage 18.97, it also recorded complete death of growth for breast cancer cell line MDA-MB-231/ATCC with growth percentage −1.21 and moderate anticancer activities for leukaemia cell line CCRF-CEM, CNS cancer cell line SNB- 75, renal cancer cell line UO-31 and breast cancer cell line HS 578 T with growth percentage from 35.46 to 47.83 ( Supplementary Table S4 , .

    Techniques:

    Effect of compounds on cancer cell viability at 100 µM concentration after 72 h of incubation against human triple-negative breast cancer MDA-MB-231 and human pancreatic carcinoma Panc-1 cell lines by MTT assay, n = 3.

    Journal: International Journal of Molecular Sciences

    Article Title: The Effect of 4-(Dimethylamino)phenyl-5-oxopyrrolidines on Breast and Pancreatic Cancer Cell Colony Formation, Migration, and Growth of Tumor Spheroids

    doi: 10.3390/ijms25031834

    Figure Lengend Snippet: Effect of compounds on cancer cell viability at 100 µM concentration after 72 h of incubation against human triple-negative breast cancer MDA-MB-231 and human pancreatic carcinoma Panc-1 cell lines by MTT assay, n = 3.

    Article Snippet: The human triple-negative breast cancer MDA-MB-231 cell line and human pancreatic carcinoma cell line Panc-1 were obtained from the American Type Culture Collection (ATCC, Manassas, VA, USA).

    Techniques: Concentration Assay, Incubation, MTT Assay

    EC 50 values of the most active compounds 3c , 3d , 5k , and 5l obtained by MTT assay ( A ) against triple-negative breast cancer MDA-MB231, pancreatic cancer Panc-1 cell lines, and human fibroblasts (HF) after 72 h of incubation. Dose–response curves for the most active compound 5k after 72 h of incubation with MDA-MB-231, Panc-1, and HF cells ( B ). Data points are experimental values (averages of three repeats), while the lines are the fit of the standard inhibition model with the Hill coefficient of 2.5. n = 3.

    Journal: International Journal of Molecular Sciences

    Article Title: The Effect of 4-(Dimethylamino)phenyl-5-oxopyrrolidines on Breast and Pancreatic Cancer Cell Colony Formation, Migration, and Growth of Tumor Spheroids

    doi: 10.3390/ijms25031834

    Figure Lengend Snippet: EC 50 values of the most active compounds 3c , 3d , 5k , and 5l obtained by MTT assay ( A ) against triple-negative breast cancer MDA-MB231, pancreatic cancer Panc-1 cell lines, and human fibroblasts (HF) after 72 h of incubation. Dose–response curves for the most active compound 5k after 72 h of incubation with MDA-MB-231, Panc-1, and HF cells ( B ). Data points are experimental values (averages of three repeats), while the lines are the fit of the standard inhibition model with the Hill coefficient of 2.5. n = 3.

    Article Snippet: The human triple-negative breast cancer MDA-MB-231 cell line and human pancreatic carcinoma cell line Panc-1 were obtained from the American Type Culture Collection (ATCC, Manassas, VA, USA).

    Techniques: MTT Assay, Incubation, Inhibition

    Effect of compounds 3c , 3d , 5k , and 5l on cell colony formation ( A ) and growth ( B ) of the human triple-negative breast cancer MDA-MB-231 cell line and cell colony formation ( C ) and growth ( D ) of the human pancreatic ductal carcinoma cell line, n = 3. Asterisks (*) indicate p < 0.05 compared with the control (untreated cells).

    Journal: International Journal of Molecular Sciences

    Article Title: The Effect of 4-(Dimethylamino)phenyl-5-oxopyrrolidines on Breast and Pancreatic Cancer Cell Colony Formation, Migration, and Growth of Tumor Spheroids

    doi: 10.3390/ijms25031834

    Figure Lengend Snippet: Effect of compounds 3c , 3d , 5k , and 5l on cell colony formation ( A ) and growth ( B ) of the human triple-negative breast cancer MDA-MB-231 cell line and cell colony formation ( C ) and growth ( D ) of the human pancreatic ductal carcinoma cell line, n = 3. Asterisks (*) indicate p < 0.05 compared with the control (untreated cells).

    Article Snippet: The human triple-negative breast cancer MDA-MB-231 cell line and human pancreatic carcinoma cell line Panc-1 were obtained from the American Type Culture Collection (ATCC, Manassas, VA, USA).

    Techniques:

    Effect of compounds 3c , 3d , 5k , and 5l on human triple-negative MDA-MB-231 and human pancreatic Panc-1 cell migration determined by ‘wound-healing’ assay. Photos of the ‘wound’ area (marked in yellow) in the MDA-MB-231 ( A ) and Panc-1 ( D ) monolayer at the beginning and the end of the experiment. The ‘wound’ area of MDA-MB-231 cells treated with 1 µM ( B ) and 2 µM ( C ) of the tested compounds at the end of the experiment, n = 3. The ‘wound’ area of Panc-1 cells treated with 1 µM ( E ) and 2 µM ( F ) of the tested compounds at the end of the experiment, n = 3. The scale bar indicates 100 µm. Asterisks (*) indicate p < 0.05 compared with the control (untreated cells).

    Journal: International Journal of Molecular Sciences

    Article Title: The Effect of 4-(Dimethylamino)phenyl-5-oxopyrrolidines on Breast and Pancreatic Cancer Cell Colony Formation, Migration, and Growth of Tumor Spheroids

    doi: 10.3390/ijms25031834

    Figure Lengend Snippet: Effect of compounds 3c , 3d , 5k , and 5l on human triple-negative MDA-MB-231 and human pancreatic Panc-1 cell migration determined by ‘wound-healing’ assay. Photos of the ‘wound’ area (marked in yellow) in the MDA-MB-231 ( A ) and Panc-1 ( D ) monolayer at the beginning and the end of the experiment. The ‘wound’ area of MDA-MB-231 cells treated with 1 µM ( B ) and 2 µM ( C ) of the tested compounds at the end of the experiment, n = 3. The ‘wound’ area of Panc-1 cells treated with 1 µM ( E ) and 2 µM ( F ) of the tested compounds at the end of the experiment, n = 3. The scale bar indicates 100 µm. Asterisks (*) indicate p < 0.05 compared with the control (untreated cells).

    Article Snippet: The human triple-negative breast cancer MDA-MB-231 cell line and human pancreatic carcinoma cell line Panc-1 were obtained from the American Type Culture Collection (ATCC, Manassas, VA, USA).

    Techniques: Migration, Wound Healing Assay

    Effect of compounds 3c , 3d , 5k , and 5l on 3D cell cultures. ( A ) Human pancreatic carcinoma Panc-1 spheroid size (brighter green color) and viability of cells in spheroids (lighter green color) at the end of the experiment. ( B ) Human triple-negative breast cancer MDA-MB-231 spheroid size (brighter blue color) and viability of cells in spheroids (lighter blue color) at the end of the experiment. ( C ) Photos of Panc-1 tumor spheroids at the beginning (Day 0), in the middle (Day 4), and at the end (Day 8) of the experiment (after incubation with 10 µM of the compounds). ( D ) Photos of MDA-MB-231 tumor spheroids at the beginning (Day 0), in the middle (Day 4), and at the end (Day 8) of the experiment (after incubation with 10 µM of the compounds). Asterisks (*) indicate p < 0.05 compared with the control (untreated spheroids); crosses (×) indicate means; inner dashes indicate medians; and whiskers indicate maximum and minimum values. Scale bars indicate 200 µm.

    Journal: International Journal of Molecular Sciences

    Article Title: The Effect of 4-(Dimethylamino)phenyl-5-oxopyrrolidines on Breast and Pancreatic Cancer Cell Colony Formation, Migration, and Growth of Tumor Spheroids

    doi: 10.3390/ijms25031834

    Figure Lengend Snippet: Effect of compounds 3c , 3d , 5k , and 5l on 3D cell cultures. ( A ) Human pancreatic carcinoma Panc-1 spheroid size (brighter green color) and viability of cells in spheroids (lighter green color) at the end of the experiment. ( B ) Human triple-negative breast cancer MDA-MB-231 spheroid size (brighter blue color) and viability of cells in spheroids (lighter blue color) at the end of the experiment. ( C ) Photos of Panc-1 tumor spheroids at the beginning (Day 0), in the middle (Day 4), and at the end (Day 8) of the experiment (after incubation with 10 µM of the compounds). ( D ) Photos of MDA-MB-231 tumor spheroids at the beginning (Day 0), in the middle (Day 4), and at the end (Day 8) of the experiment (after incubation with 10 µM of the compounds). Asterisks (*) indicate p < 0.05 compared with the control (untreated spheroids); crosses (×) indicate means; inner dashes indicate medians; and whiskers indicate maximum and minimum values. Scale bars indicate 200 µm.

    Article Snippet: The human triple-negative breast cancer MDA-MB-231 cell line and human pancreatic carcinoma cell line Panc-1 were obtained from the American Type Culture Collection (ATCC, Manassas, VA, USA).

    Techniques: Incubation

    Cell uptake of [89Zr]Zr-DFO-NY003 and [177Lu]Lu-DTPA-NY003 in MDA-MB-231 ( A ) and ACHN ( B ) cells at different time points. (Block = co-incubation with non-radiolabeled NY003 and radiotracers for 48 and 72 h. Values were expressed as mean ± SD, n = 6. **Means P < 0.01, ns means not statistically significant)

    Journal: EJNMMI Radiopharmacy and Chemistry

    Article Title: Preclinical evaluation of the theranostic potential of 89 Zr/ 177 Lu-labeled anti-TROP-2 antibody in triple-negative breast cancer model

    doi: 10.1186/s41181-023-00235-x

    Figure Lengend Snippet: Cell uptake of [89Zr]Zr-DFO-NY003 and [177Lu]Lu-DTPA-NY003 in MDA-MB-231 ( A ) and ACHN ( B ) cells at different time points. (Block = co-incubation with non-radiolabeled NY003 and radiotracers for 48 and 72 h. Values were expressed as mean ± SD, n = 6. **Means P < 0.01, ns means not statistically significant)

    Article Snippet: Triple-negative breast cancer cell line MDA-MB-231 cell line (TROP-2 mild expression) was obtained from American Type Culture Collection (ATCC) and cultured in DMEM medium (Gibco) supplemented with 10% (v/v) fetal calf serum (Thermo Fisher), 2 mM L-Glutamine (Gibco), 200 μg/mL GENETICIN (Gibco), 100 U/mL Penicillin, and 100 μg/mL Streptomycin (Hyclone).

    Techniques: Blocking Assay, Incubation

    The representative images of TROP-2 IHC staining of TROP-2 positive MDA-MB-231 ( A ) and TROP-2 negative ACHN ( B ) xenograft tumors, illustrating the weak to moderate specific TROP-2 over-expression of MDA-MB-231 tumor samples

    Journal: EJNMMI Radiopharmacy and Chemistry

    Article Title: Preclinical evaluation of the theranostic potential of 89 Zr/ 177 Lu-labeled anti-TROP-2 antibody in triple-negative breast cancer model

    doi: 10.1186/s41181-023-00235-x

    Figure Lengend Snippet: The representative images of TROP-2 IHC staining of TROP-2 positive MDA-MB-231 ( A ) and TROP-2 negative ACHN ( B ) xenograft tumors, illustrating the weak to moderate specific TROP-2 over-expression of MDA-MB-231 tumor samples

    Article Snippet: Triple-negative breast cancer cell line MDA-MB-231 cell line (TROP-2 mild expression) was obtained from American Type Culture Collection (ATCC) and cultured in DMEM medium (Gibco) supplemented with 10% (v/v) fetal calf serum (Thermo Fisher), 2 mM L-Glutamine (Gibco), 200 μg/mL GENETICIN (Gibco), 100 U/mL Penicillin, and 100 μg/mL Streptomycin (Hyclone).

    Techniques: Immunohistochemistry, Over Expression

    Micro-PET images in MDA-MB-231 tumor-bearing mice at different time points. A Representative MIP images of [ 89 Zr]Zr-DFO-NY003 (5.55–7.4 MBq) in MDA-MB-231 model at different time points p.i.. B Representative MIP images of [ 89 Zr]Zr-DFO-IgG (5.55 ~ 7.4 MBq) in the MDA-MB-231 model at different time points p.i.. C Representative MIP images of co-injection of [ 89 Zr]Zr-DFO-NY003 (5.55 ~ 7.4 MBq) and cold NY003 in MDA-MB-231 model at different time points p.i.. D–G Quantitative ROI analysis showed that the tumor uptake of [ 89 Zr]Zr-DFO-NY003 in MDA-MB-231 ( n = 3) was significantly higher than that of blocking and [ 89 Zr]Zr-DFO-IgG groups ( P < 0.001). The radioactive uptake of the heart (blood), liver, and kidney gradually decreased within the time tested

    Journal: EJNMMI Radiopharmacy and Chemistry

    Article Title: Preclinical evaluation of the theranostic potential of 89 Zr/ 177 Lu-labeled anti-TROP-2 antibody in triple-negative breast cancer model

    doi: 10.1186/s41181-023-00235-x

    Figure Lengend Snippet: Micro-PET images in MDA-MB-231 tumor-bearing mice at different time points. A Representative MIP images of [ 89 Zr]Zr-DFO-NY003 (5.55–7.4 MBq) in MDA-MB-231 model at different time points p.i.. B Representative MIP images of [ 89 Zr]Zr-DFO-IgG (5.55 ~ 7.4 MBq) in the MDA-MB-231 model at different time points p.i.. C Representative MIP images of co-injection of [ 89 Zr]Zr-DFO-NY003 (5.55 ~ 7.4 MBq) and cold NY003 in MDA-MB-231 model at different time points p.i.. D–G Quantitative ROI analysis showed that the tumor uptake of [ 89 Zr]Zr-DFO-NY003 in MDA-MB-231 ( n = 3) was significantly higher than that of blocking and [ 89 Zr]Zr-DFO-IgG groups ( P < 0.001). The radioactive uptake of the heart (blood), liver, and kidney gradually decreased within the time tested

    Article Snippet: Triple-negative breast cancer cell line MDA-MB-231 cell line (TROP-2 mild expression) was obtained from American Type Culture Collection (ATCC) and cultured in DMEM medium (Gibco) supplemented with 10% (v/v) fetal calf serum (Thermo Fisher), 2 mM L-Glutamine (Gibco), 200 μg/mL GENETICIN (Gibco), 100 U/mL Penicillin, and 100 μg/mL Streptomycin (Hyclone).

    Techniques: Micro-PET, Injection, Blocking Assay

    SPECT images in MDA-MB-231 and ACHN tumor-bearing mice at different time points p.i.. SPECT images showed specific high [ 177 Lu]Lu-DTPA-NY003 accumulation in tumors which can be blocked by cold NY003

    Journal: EJNMMI Radiopharmacy and Chemistry

    Article Title: Preclinical evaluation of the theranostic potential of 89 Zr/ 177 Lu-labeled anti-TROP-2 antibody in triple-negative breast cancer model

    doi: 10.1186/s41181-023-00235-x

    Figure Lengend Snippet: SPECT images in MDA-MB-231 and ACHN tumor-bearing mice at different time points p.i.. SPECT images showed specific high [ 177 Lu]Lu-DTPA-NY003 accumulation in tumors which can be blocked by cold NY003

    Article Snippet: Triple-negative breast cancer cell line MDA-MB-231 cell line (TROP-2 mild expression) was obtained from American Type Culture Collection (ATCC) and cultured in DMEM medium (Gibco) supplemented with 10% (v/v) fetal calf serum (Thermo Fisher), 2 mM L-Glutamine (Gibco), 200 μg/mL GENETICIN (Gibco), 100 U/mL Penicillin, and 100 μg/mL Streptomycin (Hyclone).

    Techniques: Single Photon Emission Computed Tomography

    LOX inhibition impairs osteoclast differentiation induced by human MDA-MB 231 and Hs 578T breast cancer cells. In vitro osteoclast differentiation of murine bone marrow cells treated with M-CSF + RANKL in combination with the conditioned medium from MDA-MB 231 or Hs 568T Ctrl, LOX(−) or LOXL2(−) cells. (A) TRAP staining of differentiated osteoclasts at day 7 of culture. Images shown are examples that best illustrate data obtained for each group. (B) Mature osteoclasts were quantified as multinucleated (more than 3 nuclei) TRAP-positive cells (n = 4). (C) Quantitative PCR determination of the relative levels of Rank , Trap , integrin β 3 ( itgb3 ) and cathepsin K ( cathK ) in murine bone marrow cells treated for 7 days with M-CSF + RANKL and the conditioned medium from MDA-MB 231 Ctrl, LOX(−), or LOXL2(−) cells (n = 9). Relative gene expression levels were normalized according to the C t value of the gene encoding the mouse ribosomal protein L32 *, **, ***: p < 0.05, 0.01 and 0.001, respectively.

    Journal: Journal of Bone Oncology

    Article Title: LOX, but not LOXL2, promotes bone metastasis formation and bone destruction in triple-negative breast cancer

    doi: 10.1016/j.jbo.2024.100522

    Figure Lengend Snippet: LOX inhibition impairs osteoclast differentiation induced by human MDA-MB 231 and Hs 578T breast cancer cells. In vitro osteoclast differentiation of murine bone marrow cells treated with M-CSF + RANKL in combination with the conditioned medium from MDA-MB 231 or Hs 568T Ctrl, LOX(−) or LOXL2(−) cells. (A) TRAP staining of differentiated osteoclasts at day 7 of culture. Images shown are examples that best illustrate data obtained for each group. (B) Mature osteoclasts were quantified as multinucleated (more than 3 nuclei) TRAP-positive cells (n = 4). (C) Quantitative PCR determination of the relative levels of Rank , Trap , integrin β 3 ( itgb3 ) and cathepsin K ( cathK ) in murine bone marrow cells treated for 7 days with M-CSF + RANKL and the conditioned medium from MDA-MB 231 Ctrl, LOX(−), or LOXL2(−) cells (n = 9). Relative gene expression levels were normalized according to the C t value of the gene encoding the mouse ribosomal protein L32 *, **, ***: p < 0.05, 0.01 and 0.001, respectively.

    Article Snippet: Human Hs578T and MDA-MB-231 breast cancer cell lines were obtained from the American Type Culture Collection (Manassas, VA) and authenticated using short tandem repeat analysis.

    Techniques: Inhibition, In Vitro, Staining, Real-time Polymerase Chain Reaction, Expressing

    LOX overexpression in breast cancer cells increases the production of the pro-osteoclastic cytokine IL-6. (A) ELISA measurement of IL-6 in the conditioned medium from +/− βAPN-treated B02 LOX(+), LOX(−), LOXL2(+) or LOXL2(−) cells. Data are the mean ± SD of three independent experiments. (B) ELISA measurement of IL-6 in the conditioned medium from MDA-MB-231 Ctrl, LOX(−), or LOXL2(−) cells. Data are the mean ± SD of three independent experiments. (C) Correlation between IL-6 expression with that of LOX (left-hand panel) or LOXL2 (right-hand panel) in ER-negative primary mammary tumors (n = 78) from a cohort of breast cancer patients (GSE2034). Pearson correlation coefficients and P-values obtained with the PRISM 9 software are indicated. **, ***: p < 0.01 and 0.001, respectively.

    Journal: Journal of Bone Oncology

    Article Title: LOX, but not LOXL2, promotes bone metastasis formation and bone destruction in triple-negative breast cancer

    doi: 10.1016/j.jbo.2024.100522

    Figure Lengend Snippet: LOX overexpression in breast cancer cells increases the production of the pro-osteoclastic cytokine IL-6. (A) ELISA measurement of IL-6 in the conditioned medium from +/− βAPN-treated B02 LOX(+), LOX(−), LOXL2(+) or LOXL2(−) cells. Data are the mean ± SD of three independent experiments. (B) ELISA measurement of IL-6 in the conditioned medium from MDA-MB-231 Ctrl, LOX(−), or LOXL2(−) cells. Data are the mean ± SD of three independent experiments. (C) Correlation between IL-6 expression with that of LOX (left-hand panel) or LOXL2 (right-hand panel) in ER-negative primary mammary tumors (n = 78) from a cohort of breast cancer patients (GSE2034). Pearson correlation coefficients and P-values obtained with the PRISM 9 software are indicated. **, ***: p < 0.01 and 0.001, respectively.

    Article Snippet: Human Hs578T and MDA-MB-231 breast cancer cell lines were obtained from the American Type Culture Collection (Manassas, VA) and authenticated using short tandem repeat analysis.

    Techniques: Over Expression, Enzyme-linked Immunosorbent Assay, Expressing, Software