Journal: The EMBO Journal

Article Title: Genome‐wide cooperation of EMT transcription factor ZEB 1 with YAP and AP ‐1 in breast cancer

doi: 10.15252/embj.2019103209

Figure Lengend Snippet: Immunofluorescence staining of ZEB1, the AP‐1 factors JUN and FOSL1, and the Hippo pathway effector TAZ in MDA‐MB‐231 cells. All four proteins show a clear nuclear localisation. Scale bar = 20 μm. In situ proximity ligation assay (PLA) of ZEB1 with JUN, FOSL1 and TAZ shows close proximity of ZEB1 with JUN and FOSL1 but not TAZ in the nucleus of BT549 and Hs578T breast cancer cells indicated by red fluorescent dots. In the upper panel, representative microscopic images are shown. Scale bar = 20 μm. The lower panel shows a quantification of the PLA shown above. n = 2–3; shown is one representative experiment with means ± s.e.m. of at least 900 cells per condition; *** P ≤ 0.001; 2‐way ANOVA plus Tukey′s post‐test.

Article Snippet: MDA‐MB‐231, MCF7, BT549, Hs578T and MCF10A cells were purchased from the American Type Culture Collection (ATCC).

Techniques: Immunofluorescence, Staining, In Situ, Proximity Ligation Assay

Journal: The EMBO Journal

Article Title: Genome‐wide cooperation of EMT transcription factor ZEB 1 with YAP and AP ‐1 in breast cancer

doi: 10.15252/embj.2019103209

Figure Lengend Snippet: qRT–PCR of ZEB1, FOSL1 and YAP, 11 ZEB1/YAP/JUN‐activated target genes and one ZEB1‐only repressed target gene, CDH1, in MCF10A cells after 7 days of TGFβ treatment compared to control treated cells. Both, control and TGFß‐treated cells were treated with control siRNA for 72 h before harvest. n = 3; shown is mean ± s.e.m. Luciferase reporter assay in BT549 and Hs578T cells comparing control and transient siRNA‐mediated knockdown of ZEB1. Reporter constructs of regulatory regions of genes shown in Fig (B) and (C). n = 3; shown is mean ± s.e.m.; * P ≤ 0.05; ** P ≤ 0.01; *** P ≤ 0.001; unpaired two‐tailed Student's t ‐test.

Article Snippet: MDA‐MB‐231, MCF7, BT549, Hs578T and MCF10A cells were purchased from the American Type Culture Collection (ATCC).

Techniques: Quantitative RT-PCR, Control, Luciferase, Reporter Assay, Knockdown, Construct, Two Tailed Test

Journal: The EMBO Journal

Article Title: Reciprocal priming between receptor tyrosine kinases at recycling endosomes orchestrates cellular signalling outputs

doi: 10.15252/embj.2020107182

Figure Lengend Snippet:

Article Snippet: BT549 , LGC ATCC® , HTB‐122.

Techniques: Virus, Recombinant, Extraction, Multiplex sample analysis, Transfection, Molecular Weight, Marker, Reverse Transcription, Protease Inhibitor, Membrane, Cell Culture, Clinical Proteomics, Spectroscopy, Imaging, cDNA Synthesis, Plasmid Preparation, Negative Control, Mutagenesis, Clone Assay, Generated, Software, Microscopy

Journal: Scientific Reports

Article Title: TRAF6 as a potential target in advanced breast cancer: a systematic review, meta-analysis, and bioinformatics validation

doi: 10.1038/s41598-023-31557-0

Figure Lengend Snippet: Summary of meta-analysis of included studies showing significant association of in vitro breast cancer cell behaviour and modulation of TRAF2/4/6.

Article Snippet: , Proliferation , Pharmacological inhibition , BT549 (4) MCF7 (4) MDA-MB-231 (8) , − 5.78 [− 12.04, 0.47] − 9.50 [− 18.82, − 0.19] − 5.62 [− 10.34, − 0.90] , − 21.46 [− 30.40, − 12.51] , Mean Difference (IV, Random, 95% CI) , Tau 2 = 321.80; Chi 2 = 677.01, df = 15 (P < 0.00001); I 2 = 98% , Z = 4.70 (P < 0.00001).

Techniques: In Vitro, Migration, Inhibition

Journal: Scientific Reports

Article Title: TRAF6 as a potential target in advanced breast cancer: a systematic review, meta-analysis, and bioinformatics validation

doi: 10.1038/s41598-023-31557-0

Figure Lengend Snippet: Summary of meta-analysis of included studies showing significant association of in vivo tumour burden and overt metastasis with modulation of TRAF2/4/6.

Article Snippet: , Proliferation , Pharmacological inhibition , BT549 (4) MCF7 (4) MDA-MB-231 (8) , − 5.78 [− 12.04, 0.47] − 9.50 [− 18.82, − 0.19] − 5.62 [− 10.34, − 0.90] , − 21.46 [− 30.40, − 12.51] , Mean Difference (IV, Random, 95% CI) , Tau 2 = 321.80; Chi 2 = 677.01, df = 15 (P < 0.00001); I 2 = 98% , Z = 4.70 (P < 0.00001).

Techniques: In Vivo, Inhibition

Journal: Frontiers in Immunology

Article Title: HMGA2 interacts with KAT6A to regulate MMPs chromatin architecture and promote triple-negative breast cancer metastasis

doi: 10.3389/fimmu.2025.1590368

Figure Lengend Snippet: HMGA2 is highly expressed in TNBC and associates with patients’ poor prognosis. (A) HMGA2 mRNA expression levels in normal and tumor tissues. Data were obtained from GDC TCGA. (B) HMGA2 mRNA expression levels in TNBC and non-TNBC tissues. Data were obtained from GDC TCGA. Breast cancer can be classified according to their expression of estrogen receptor (ER), progesterone receptor (PR), HER2 and Ki67. (C) The representative HMGA2 IHC images of the FFPE samples, 6 cases of normal tissues, 60 cases of TNBC tissues and 20 cases of non-TNBC tissues. Scale bar: 50 μm. Magnification: 400 ×. (D, E) mRNA (D) and protein (E) of HMGA2 expression in TNBC cells (MDA-MB-231 and BT549) were higher than those of MCF7 and T-47D cell lines. (F, G) TNBC patients with high HMGA2 expression exhibited poor prognosis. TNBC patients from TMUGH. ns, no significance; **p<0.01, ***p<0.001, ****p<0.0001.

Article Snippet: MDA-MB-231 and BT549 cells were obtained from the China Center for Type Culture Collection (CCTCC) and authenticated by short tandem repeat (STR) profiling.

Techniques: Expressing

Journal: Frontiers in Immunology

Article Title: HMGA2 interacts with KAT6A to regulate MMPs chromatin architecture and promote triple-negative breast cancer metastasis

doi: 10.3389/fimmu.2025.1590368

Figure Lengend Snippet: HMGA2 interacts with KAT6A regulating the expression of MMPs. (A) Proteins from the IP assay on MDA-MB-231 cells were separated by SDS-PAGE and detected by Coomassie brilliant blue staining. (B, C) Interaction between HMGA2 and KAT6A were verified in MDA-MB-231 (B) and BT549 (C) cells by Co-IP assay. (D) Schematic of various HMGA2 truncations. (E) Total cell lysates from HEK293T cells expressing different truncations of HMGA2 were subjected to IP with antibodies against Flag tags. (F–H) ChIP results demonstrated the interaction of H3K23ac with promoters of MMPs in TNBC cells. *p<0.05, **p<0.01, ***p<0.001, ****p<0.0001.

Article Snippet: MDA-MB-231 and BT549 cells were obtained from the China Center for Type Culture Collection (CCTCC) and authenticated by short tandem repeat (STR) profiling.

Techniques: Expressing, SDS Page, Staining, Co-Immunoprecipitation Assay

Journal: Journal of Experimental & Clinical Cancer Research : CR

Article Title: circGFRA1 and GFRA1 act as ceRNAs in triple negative breast cancer by regulating miR-34a

doi: 10.1186/s13046-017-0614-1

Figure Lengend Snippet: circRNA expression profiles in TNBC. a. The cluster heat map showed the differentially expressed circRNAs over 3-fold change. Red color indicates high expression level, and green color indicates low expression level. The black arrow indicates circGFRA1. b. The scatter plot was used for assessing the variation in circRNA expression between TNBC cell lines and HME cell line. The values of x and y axes in the scatter plot were the normalized signal values of the samples (log2 scaled). The green lines are fold-change lines. c. The top 10 upregulated circRNAs were shown. d. qRT-PCR was performed to verify the expression of the top eight upregulated circRNAs. All the data are shown as the mean ± s.e.m.

Article Snippet: Three TNBC cell lines (MDA-MB-231, BT549 and MDA-MB-468) and normal mammary epithelial cell line (MCF-10A) were analyzed by Arraystar Human circRNA Array V2.

Techniques: Expressing, Quantitative RT-PCR

Journal: Journal of Experimental & Clinical Cancer Research : CR

Article Title: circGFRA1 and GFRA1 act as ceRNAs in triple negative breast cancer by regulating miR-34a

doi: 10.1186/s13046-017-0614-1

Figure Lengend Snippet: circGFRA1 is upregulated and correlated with poor clinical outcomes in TNBC. a. The expression level of circGFRA1 was determined by qRT-PCR in 11 different mammary cell lines, including two HME cell line (MCF-10A and 184A1) and nine breast cancer cell lines. circGFRA1 expression was normalized using β-actin expression. b. The expression level of circGFRA1 in 51 TNBC tissues and their matched normal adjacent tissues was determined by qRT-PCR. c. OS curves for 222 TNBC patients with high or low circGFRA1 expression. d. DFS curves for 222 TNBC patients with high or low circGFRA1 expression. All the data are shown as the mean ± s.e.m.

Article Snippet: Three TNBC cell lines (MDA-MB-231, BT549 and MDA-MB-468) and normal mammary epithelial cell line (MCF-10A) were analyzed by Arraystar Human circRNA Array V2.

Techniques: Expressing, Quantitative RT-PCR

Journal: Journal of Experimental & Clinical Cancer Research : CR

Article Title: circGFRA1 and GFRA1 act as ceRNAs in triple negative breast cancer by regulating miR-34a

doi: 10.1186/s13046-017-0614-1

Figure Lengend Snippet: circGFRA1 serves as a sponge for miR-34a. a. The levels of nuclear control transcript (U6), cytoplasmic control transcript (GAPDH mRNA) and circGFRA1 were assessed by qRT-PCR in nuclear and cytoplasmic fractions. b. The predicted binding sites of miR-34a within circGFRA1 were shown. c. Cells were transfected with miR-34a mimic or miR-34a inhibitor, qRT-PCR analysis demonstrated that the transfection was successful. d. Luciferase assay of MDA-MB-231 cells cotransfected with miR-34a mimics or miR-34a inhibitor and luciferase reporter containing circGFRA1 3′-UTR (circGFRA1 wt) or mutant construct (circGFRA1 mut). e. Cells were transfected as described, and the expression of circGFRA1 was determined by qRT-PCR. f. Cells were transfected with si-NC or si-circGFRA1, and the expression of miR-34a was determined by qRT-PCR. U6 snRNA was used as an internal control. g. MS2-based RIP assay in TNBC cells transfected with MS2bs-circGFRA1, MS2bs-circGFRA1mt, or MS2bs-Rluc (control vector) ( n = 3). All the data are shown as the mean ± s.e.m., * P < 0.05 and ** P < 0.01

Article Snippet: Three TNBC cell lines (MDA-MB-231, BT549 and MDA-MB-468) and normal mammary epithelial cell line (MCF-10A) were analyzed by Arraystar Human circRNA Array V2.

Techniques: Control, Quantitative RT-PCR, Binding Assay, Transfection, Luciferase, Mutagenesis, Construct, Expressing, Plasmid Preparation

Journal: Journal of Experimental & Clinical Cancer Research : CR

Article Title: circGFRA1 and GFRA1 act as ceRNAs in triple negative breast cancer by regulating miR-34a

doi: 10.1186/s13046-017-0614-1

Figure Lengend Snippet: circGFRA1 and GFRA1 act as ceRNAs in TNBC through regulation of miR-34a. a. The expression level of GFRA1 was determined by qRT-PCR (left) and western blot (right) in 11 mammary cell lines. β-actin was used as a control. b. The expression level of GFRA1 in 51 TNBC tissues and their matched normal adjacent tissues was determined by qRT-PCR. c. The predicted binding sites of miR-34a within GFRA1 were shown. d. Luciferase assay of MDA-MB-231 cells cotransfected with miR-34a mimics or miR-34a inhibitor and luciferase reporter containing GFRA1 3′-UTR (GFRA1 wt) or mutant construct (GFRA1 mut). e. Cells were transfected as described, and the expression of GFRA1 was determined by qRT-PCR. f. Cells were transfected as described, and the expression of GFRA1 was determined by western blot analysis. G. Cells were transfected with si-NC or si-GFRA1, and the expression of miR-34a was determined by qRT-PCR. h. Cells were transfected with si-NC, si-circGFRA1 or si-circGFRA1 + miR-34a inhibitor, and the expression of GFRA1 was determined by qRT-PCR. i. Cells were transfected with si-NC, si-GFRA1 or si-GFRA1 + miR-34a inhibitor, and the expression of circGFRA1 was determined by qRT-PCR. j. Cells were transfected with miR-NC, miR-34a or si-circGFRA1 + miR-34a, and CCK8 assay was performed to assess cell proliferation. k. Cells were transfected as described, and apoptosis assay was performed after transfection. All the data are shown as the mean ± s.e.m., * P < 0.05 and ** P < 0.01

Article Snippet: Three TNBC cell lines (MDA-MB-231, BT549 and MDA-MB-468) and normal mammary epithelial cell line (MCF-10A) were analyzed by Arraystar Human circRNA Array V2.

Techniques: Expressing, Quantitative RT-PCR, Western Blot, Control, Binding Assay, Luciferase, Mutagenesis, Construct, Transfection, CCK-8 Assay, Apoptosis Assay

Journal: iScience

Article Title: Conserved role of FOXC1 in TNBC is parallel to FOXA1 in ER+ breast cancer

doi: 10.1016/j.isci.2024.110500

Figure Lengend Snippet: Identification of FOXC1 cofactors (A) Protein co-interactors of FOXC1 identified in Hs578t and BT-549 via RIME in two biological replicates, after subtracting those identified in negative IgG control. (B) Western blot depicting co-immunoprecipitation of FOXC1 using antibody against NR2F2 with input and IgG negative controls.

Article Snippet: Bt549 , AddexBio , C0006017, RRID:CVCL_1092.

Techniques: Control, Western Blot, Immunoprecipitation

Journal: iScience

Article Title: Conserved role of FOXC1 in TNBC is parallel to FOXA1 in ER+ breast cancer

doi: 10.1016/j.isci.2024.110500

Figure Lengend Snippet:

Article Snippet: Bt549 , AddexBio , C0006017, RRID:CVCL_1092.

Techniques: Recombinant, Bicinchoninic Acid Protein Assay, Sequencing, Generated, Gene Expression, Mass Spectrometry, Knock-Out, Software, Blocking Assay