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ATCC bt 474 cells cat htb 20 cells

MCF-7 and <t>BT-474</t> cells were cultured on soft (0.5 kPa) or stiff (9 kPa) supports for 2 weeks before subsequent analysis. a Docetaxel (Doc) induced apoptosis of MCF-7 and BT-474 was assessed by flow cytometry. The proportions of Annexin V + cells are shown. b Quantitation of apoptotic cells (Annexin V + ) in MCF-7 and BT-474 cells in (a) ( n = 5 biologically independent experiments). Mean ± SD, **** p < 0.0001 by two-sided two-way ANOVA with Tukey test. c MCF-7 and BT-474 cells cultured on soft or stiff supports were treated with docetaxel or cisplatin. Cell viability was evaluated after 24 h by CCK8 assays ( n = 5 biologically independent experiments). Mean ± SD, **** p < 0.0001 by two-sided Student’s t test. d Western blots for cleaved/uncleaved caspase-3 and PARP in docetaxel-treated MCF-7 and BT-474 cells cultured on soft or stiff supports. Representative images from 3 biologically independent experiments are shown. e The percentage of CD44 high CD24 low cells in MCF-7 and BT-474 cells cultured on soft or stiff supports were determined by flow cytometry. f Quantitation of CD44 high CD24 low cells in (e) ( n = 5 biologically independent experiments). Mean ± SD, **** p < 0.0001 by two-sided Student’s t test. For b-d and f , source data are provided as a Source Data file.

Bt 474 Cells Cat Htb 20 Cells, 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 "Niche stiffness sustains cancer stemness via TAZ and NANOG phase separation"

Article Title: Niche stiffness sustains cancer stemness via TAZ and NANOG phase separation

Journal: Nature Communications

doi: 10.1038/s41467-023-35856-y

MCF-7 and BT-474 cells were cultured on soft (0.5 kPa) or stiff (9 kPa) supports for 2 weeks before subsequent analysis. a Docetaxel (Doc) induced apoptosis of MCF-7 and BT-474 was assessed by flow cytometry. The proportions of Annexin V + cells are shown. b Quantitation of apoptotic cells (Annexin V + ) in MCF-7 and BT-474 cells in (a) ( n = 5 biologically independent experiments). Mean ± SD, **** p < 0.0001 by two-sided two-way ANOVA with Tukey test. c MCF-7 and BT-474 cells cultured on soft or stiff supports were treated with docetaxel or cisplatin. Cell viability was evaluated after 24 h by CCK8 assays ( n = 5 biologically independent experiments). Mean ± SD, **** p < 0.0001 by two-sided Student’s t test. d Western blots for cleaved/uncleaved caspase-3 and PARP in docetaxel-treated MCF-7 and BT-474 cells cultured on soft or stiff supports. Representative images from 3 biologically independent experiments are shown. e The percentage of CD44 high CD24 low cells in MCF-7 and BT-474 cells cultured on soft or stiff supports were determined by flow cytometry. f Quantitation of CD44 high CD24 low cells in (e) ( n = 5 biologically independent experiments). Mean ± SD, **** p < 0.0001 by two-sided Student’s t test. For b-d and f , source data are provided as a Source Data file.

Figure Legend Snippet: MCF-7 and BT-474 cells were cultured on soft (0.5 kPa) or stiff (9 kPa) supports for 2 weeks before subsequent analysis. a Docetaxel (Doc) induced apoptosis of MCF-7 and BT-474 was assessed by flow cytometry. The proportions of Annexin V + cells are shown. b Quantitation of apoptotic cells (Annexin V + ) in MCF-7 and BT-474 cells in (a) ( n = 5 biologically independent experiments). Mean ± SD, **** p < 0.0001 by two-sided two-way ANOVA with Tukey test. c MCF-7 and BT-474 cells cultured on soft or stiff supports were treated with docetaxel or cisplatin. Cell viability was evaluated after 24 h by CCK8 assays ( n = 5 biologically independent experiments). Mean ± SD, **** p < 0.0001 by two-sided Student’s t test. d Western blots for cleaved/uncleaved caspase-3 and PARP in docetaxel-treated MCF-7 and BT-474 cells cultured on soft or stiff supports. Representative images from 3 biologically independent experiments are shown. e The percentage of CD44 high CD24 low cells in MCF-7 and BT-474 cells cultured on soft or stiff supports were determined by flow cytometry. f Quantitation of CD44 high CD24 low cells in (e) ( n = 5 biologically independent experiments). Mean ± SD, **** p < 0.0001 by two-sided Student’s t test. For b-d and f , source data are provided as a Source Data file.

Techniques Used: Cell Culture, Flow Cytometry, Quantitation Assay, Western Blot

MCF-7 and BT-474 cells were cultured on soft or stiff supports for 2 weeks before subsequent analysis. In some experiments, cells were pre-transduced with TAZ shRNA (shTAZ-1 or shTAZ-2) or GFP shRNA (shGFP). a The expression of TAZ in MCF-7 and BT-474 cells cultured on soft or stiff supports was determined by western blot ( n = 3 biologically independent experiments). b Representative immunofluorescence staining of TAZ in MCF-7 and BT-474 cells cultured on soft or stiff substrates. The insert represents a magnification from the indicated area (white square). Scale bars, 10 μm. c Proportion of cancer cells exhibiting mainly nuclear TAZ localization (N > C), diffuse distribution of TAZ in nucleus and cytoplasm (N = C), or preferential cytoplasmic TAZ (N < C or undetectable). 20 cells were assessed per group, experiments were independently repeated 4 times. Mean ± SD, **** p < 0.0001 by two-tailed chi-square test. d Docetaxel induced apoptosis of MCF-7 and BT-474 was assessed by flow cytometry. See Supplementary Fig. for quantification. e MCF-7 and BT-474 cells cultured on soft or stiff supports were treated with docetaxel or cisplatin. Cell viability was evaluated by CCK8 assay ( n = 5 biologically independent experiments). Mean ± SD, **** p < 0.0001 by two-sided one-way ANOVA with Tukey test. f Representative images of CD44 high CD24 low cells in MCF-7 and BT-474 cells were detected by flow cytometry. See Supplementary Fig. for quantification. g Western blots of TAZ, SOX2, and OCT4 in MCF-7 and BT-474 cells cultured on soft or stiff supports ( n = 3 biologically independent experiments). For data presented in the same figure panel, the samples were derived from the same experiment and blots were processed in parallel. For a , c , e and g , source data are provided as a Source Data file.

Figure Legend Snippet: MCF-7 and BT-474 cells were cultured on soft or stiff supports for 2 weeks before subsequent analysis. In some experiments, cells were pre-transduced with TAZ shRNA (shTAZ-1 or shTAZ-2) or GFP shRNA (shGFP). a The expression of TAZ in MCF-7 and BT-474 cells cultured on soft or stiff supports was determined by western blot ( n = 3 biologically independent experiments). b Representative immunofluorescence staining of TAZ in MCF-7 and BT-474 cells cultured on soft or stiff substrates. The insert represents a magnification from the indicated area (white square). Scale bars, 10 μm. c Proportion of cancer cells exhibiting mainly nuclear TAZ localization (N > C), diffuse distribution of TAZ in nucleus and cytoplasm (N = C), or preferential cytoplasmic TAZ (N < C or undetectable). 20 cells were assessed per group, experiments were independently repeated 4 times. Mean ± SD, **** p < 0.0001 by two-tailed chi-square test. d Docetaxel induced apoptosis of MCF-7 and BT-474 was assessed by flow cytometry. See Supplementary Fig. for quantification. e MCF-7 and BT-474 cells cultured on soft or stiff supports were treated with docetaxel or cisplatin. Cell viability was evaluated by CCK8 assay ( n = 5 biologically independent experiments). Mean ± SD, **** p < 0.0001 by two-sided one-way ANOVA with Tukey test. f Representative images of CD44 high CD24 low cells in MCF-7 and BT-474 cells were detected by flow cytometry. See Supplementary Fig. for quantification. g Western blots of TAZ, SOX2, and OCT4 in MCF-7 and BT-474 cells cultured on soft or stiff supports ( n = 3 biologically independent experiments). For data presented in the same figure panel, the samples were derived from the same experiment and blots were processed in parallel. For a , c , e and g , source data are provided as a Source Data file.

Techniques Used: Cell Culture, Transduction, shRNA, Expressing, Western Blot, Immunofluorescence, Staining, Two Tailed Test, Flow Cytometry, CCK-8 Assay, Derivative Assay

a – d TAZ and NANOG were knockout or overexpressed by lentiviral transfection in MCF-7 and BT-474 cells. NANOG, TAZ, SOX2 or/and OCT4 were overexpressed in TAZ or NANOG knockout cells. Transfected cells were cultured on stiff hydrogels for 2 weeks. OE, overexpression. a (Left) Representative flow cytometric plot indicating the percentages of CD44 high CD24 low cells in MCF-7 and BT-474 cells. (Right) Qualification of five biologically independent experiments. **** p < 0.0001; for MCF-7 cells, *** p = 0.0003, ** p = 0.0039, ns=0.9899; for BT-474 cells, *** p = 0.0008, ns=0.9982. b Cell viability of MCF-7 and BT-474 cells treated with various concentrations of docetaxel and cisplatin for 24 h was determined by CCK8 assays ( n = 4 biologically independent experiments). P values compared with sgTAZ + Vector group were shown in the figure. c (Left) Representative flow cytometric plot indicating the percentages of CD44 high CD24 low cells in MCF-7 and BT-474 cells. (Right) Qualification of five independent experiments. **** p < 0.0001; for MCF-7 cells, *** p = 0.0004, ** p = 0.0033, ns=0.9400; for BT-474 cells, *** p = 0.0002, ** p = 0.0013, ns=0.9982. d Cell viability of MCF-7 and BT-474 cells treated with various concentrations of docetaxel and cisplatin for 24 h was determined by CCK8 assays ( n = 4 biologically independent experiments). P values compared with sgNANOG + Vector group were shown in the figure. For panels (a– d) , data are mean ± SD; p values were calculated by two-sided one-way ANOVA with Tukey test. e , f MCF-7 cells were cultured on soft or stiff hydrogels for 2 weeks. Then ChIP was performed using cell lysates with indicated antibodies. e The precipitated DNA using IgG and TAZ antibody, respectively, was quantified by qPCR with primers specific to promoter regions of Sox2 and Pou5f1 . f The precipitated DNA using IgG and NANOG antibody, respectively, was quantified by qPCR with primers specific to promoter regions of Sox2 and Pou5f1 . e , f Fold enrichment was normalized to IgG-ChIP negative control ( n = 4 biologically independent experiments). Numeric values denote mean ± SD; **** p < 0.0001 by two-sided two-way ANOVA with Tukey test. For a–f , source data are provided as a Source Data file.

Figure Legend Snippet: a – d TAZ and NANOG were knockout or overexpressed by lentiviral transfection in MCF-7 and BT-474 cells. NANOG, TAZ, SOX2 or/and OCT4 were overexpressed in TAZ or NANOG knockout cells. Transfected cells were cultured on stiff hydrogels for 2 weeks. OE, overexpression. a (Left) Representative flow cytometric plot indicating the percentages of CD44 high CD24 low cells in MCF-7 and BT-474 cells. (Right) Qualification of five biologically independent experiments. **** p < 0.0001; for MCF-7 cells, *** p = 0.0003, ** p = 0.0039, ns=0.9899; for BT-474 cells, *** p = 0.0008, ns=0.9982. b Cell viability of MCF-7 and BT-474 cells treated with various concentrations of docetaxel and cisplatin for 24 h was determined by CCK8 assays ( n = 4 biologically independent experiments). P values compared with sgTAZ + Vector group were shown in the figure. c (Left) Representative flow cytometric plot indicating the percentages of CD44 high CD24 low cells in MCF-7 and BT-474 cells. (Right) Qualification of five independent experiments. **** p < 0.0001; for MCF-7 cells, *** p = 0.0004, ** p = 0.0033, ns=0.9400; for BT-474 cells, *** p = 0.0002, ** p = 0.0013, ns=0.9982. d Cell viability of MCF-7 and BT-474 cells treated with various concentrations of docetaxel and cisplatin for 24 h was determined by CCK8 assays ( n = 4 biologically independent experiments). P values compared with sgNANOG + Vector group were shown in the figure. For panels (a– d) , data are mean ± SD; p values were calculated by two-sided one-way ANOVA with Tukey test. e , f MCF-7 cells were cultured on soft or stiff hydrogels for 2 weeks. Then ChIP was performed using cell lysates with indicated antibodies. e The precipitated DNA using IgG and TAZ antibody, respectively, was quantified by qPCR with primers specific to promoter regions of Sox2 and Pou5f1 . f The precipitated DNA using IgG and NANOG antibody, respectively, was quantified by qPCR with primers specific to promoter regions of Sox2 and Pou5f1 . e , f Fold enrichment was normalized to IgG-ChIP negative control ( n = 4 biologically independent experiments). Numeric values denote mean ± SD; **** p < 0.0001 by two-sided two-way ANOVA with Tukey test. For a–f , source data are provided as a Source Data file.

Techniques Used: Knock-Out, Transfection, Cell Culture, Over Expression, Plasmid Preparation, Negative Control

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bt 474 cells cat htb 20 cells (ATCC)

ATCC bt 474 cells cat htb 20 cells

bt 474 cells cat htb 20 cells - by Bioz Stars, 2023-03

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Journal: Nature Communications

Article Title: Niche stiffness sustains cancer stemness via TAZ and NANOG phase separation

doi: 10.1038/s41467-023-35856-y

Figure Lengend Snippet: MCF-7 and BT-474 cells were cultured on soft (0.5 kPa) or stiff (9 kPa) supports for 2 weeks before subsequent analysis. a Docetaxel (Doc) induced apoptosis of MCF-7 and BT-474 was assessed by flow cytometry. The proportions of Annexin V + cells are shown. b Quantitation of apoptotic cells (Annexin V + ) in MCF-7 and BT-474 cells in (a) ( n = 5 biologically independent experiments). Mean ± SD, **** p < 0.0001 by two-sided two-way ANOVA with Tukey test. c MCF-7 and BT-474 cells cultured on soft or stiff supports were treated with docetaxel or cisplatin. Cell viability was evaluated after 24 h by CCK8 assays ( n = 5 biologically independent experiments). Mean ± SD, **** p < 0.0001 by two-sided Student’s t test. d Western blots for cleaved/uncleaved caspase-3 and PARP in docetaxel-treated MCF-7 and BT-474 cells cultured on soft or stiff supports. Representative images from 3 biologically independent experiments are shown. e The percentage of CD44 high CD24 low cells in MCF-7 and BT-474 cells cultured on soft or stiff supports were determined by flow cytometry. f Quantitation of CD44 high CD24 low cells in (e) ( n = 5 biologically independent experiments). Mean ± SD, **** p < 0.0001 by two-sided Student’s t test. For b-d and f , source data are provided as a Source Data file.

Article Snippet: MCF-7 cells (cat#HTB-22) and BT-474 cells (cat#HTB-20) cells were obtained from American Type Culture Collection (ATCC) and cultured according to standard protocols.

Techniques: Cell Culture, Flow Cytometry, Quantitation Assay, Western Blot

Journal: Nature Communications

Article Title: Niche stiffness sustains cancer stemness via TAZ and NANOG phase separation

doi: 10.1038/s41467-023-35856-y

Figure Lengend Snippet: MCF-7 and BT-474 cells were cultured on soft or stiff supports for 2 weeks before subsequent analysis. In some experiments, cells were pre-transduced with TAZ shRNA (shTAZ-1 or shTAZ-2) or GFP shRNA (shGFP). a The expression of TAZ in MCF-7 and BT-474 cells cultured on soft or stiff supports was determined by western blot ( n = 3 biologically independent experiments). b Representative immunofluorescence staining of TAZ in MCF-7 and BT-474 cells cultured on soft or stiff substrates. The insert represents a magnification from the indicated area (white square). Scale bars, 10 μm. c Proportion of cancer cells exhibiting mainly nuclear TAZ localization (N > C), diffuse distribution of TAZ in nucleus and cytoplasm (N = C), or preferential cytoplasmic TAZ (N < C or undetectable). 20 cells were assessed per group, experiments were independently repeated 4 times. Mean ± SD, **** p < 0.0001 by two-tailed chi-square test. d Docetaxel induced apoptosis of MCF-7 and BT-474 was assessed by flow cytometry. See Supplementary Fig. for quantification. e MCF-7 and BT-474 cells cultured on soft or stiff supports were treated with docetaxel or cisplatin. Cell viability was evaluated by CCK8 assay ( n = 5 biologically independent experiments). Mean ± SD, **** p < 0.0001 by two-sided one-way ANOVA with Tukey test. f Representative images of CD44 high CD24 low cells in MCF-7 and BT-474 cells were detected by flow cytometry. See Supplementary Fig. for quantification. g Western blots of TAZ, SOX2, and OCT4 in MCF-7 and BT-474 cells cultured on soft or stiff supports ( n = 3 biologically independent experiments). For data presented in the same figure panel, the samples were derived from the same experiment and blots were processed in parallel. For a , c , e and g , source data are provided as a Source Data file.

Article Snippet: MCF-7 cells (cat#HTB-22) and BT-474 cells (cat#HTB-20) cells were obtained from American Type Culture Collection (ATCC) and cultured according to standard protocols.

Techniques: Cell Culture, Transduction, shRNA, Expressing, Western Blot, Immunofluorescence, Staining, Two Tailed Test, Flow Cytometry, CCK-8 Assay, Derivative Assay

Journal: Nature Communications

Article Title: Niche stiffness sustains cancer stemness via TAZ and NANOG phase separation

doi: 10.1038/s41467-023-35856-y

Figure Lengend Snippet: a – d TAZ and NANOG were knockout or overexpressed by lentiviral transfection in MCF-7 and BT-474 cells. NANOG, TAZ, SOX2 or/and OCT4 were overexpressed in TAZ or NANOG knockout cells. Transfected cells were cultured on stiff hydrogels for 2 weeks. OE, overexpression. a (Left) Representative flow cytometric plot indicating the percentages of CD44 high CD24 low cells in MCF-7 and BT-474 cells. (Right) Qualification of five biologically independent experiments. **** p < 0.0001; for MCF-7 cells, *** p = 0.0003, ** p = 0.0039, ns=0.9899; for BT-474 cells, *** p = 0.0008, ns=0.9982. b Cell viability of MCF-7 and BT-474 cells treated with various concentrations of docetaxel and cisplatin for 24 h was determined by CCK8 assays ( n = 4 biologically independent experiments). P values compared with sgTAZ + Vector group were shown in the figure. c (Left) Representative flow cytometric plot indicating the percentages of CD44 high CD24 low cells in MCF-7 and BT-474 cells. (Right) Qualification of five independent experiments. **** p < 0.0001; for MCF-7 cells, *** p = 0.0004, ** p = 0.0033, ns=0.9400; for BT-474 cells, *** p = 0.0002, ** p = 0.0013, ns=0.9982. d Cell viability of MCF-7 and BT-474 cells treated with various concentrations of docetaxel and cisplatin for 24 h was determined by CCK8 assays ( n = 4 biologically independent experiments). P values compared with sgNANOG + Vector group were shown in the figure. For panels (a– d) , data are mean ± SD; p values were calculated by two-sided one-way ANOVA with Tukey test. e , f MCF-7 cells were cultured on soft or stiff hydrogels for 2 weeks. Then ChIP was performed using cell lysates with indicated antibodies. e The precipitated DNA using IgG and TAZ antibody, respectively, was quantified by qPCR with primers specific to promoter regions of Sox2 and Pou5f1 . f The precipitated DNA using IgG and NANOG antibody, respectively, was quantified by qPCR with primers specific to promoter regions of Sox2 and Pou5f1 . e , f Fold enrichment was normalized to IgG-ChIP negative control ( n = 4 biologically independent experiments). Numeric values denote mean ± SD; **** p < 0.0001 by two-sided two-way ANOVA with Tukey test. For a–f , source data are provided as a Source Data file.

Article Snippet: MCF-7 cells (cat#HTB-22) and BT-474 cells (cat#HTB-20) cells were obtained from American Type Culture Collection (ATCC) and cultured according to standard protocols.

Techniques: Knock-Out, Transfection, Cell Culture, Over Expression, Plasmid Preparation, Negative Control