human breast cancer cells  (ATCC)

Journal: PLOS One

Article Title: Inhibition of HER2 signaling and breast cancer cell growth with a novel antibody targeting HER2 ECD III/IV

doi: 10.1371/journal.pone.0338127

Figure Lengend Snippet: A. Viability of BT-474 cells following 4 d treatment with a 1:3 serial dilution (0-100 nM) of the indicated antibodies, as assessed by CCK-8 assay. B. Dose-dependent inhibition of SK-BR-3 and MCF7 cell proliferation after 4 d of exposure to antibody dilutions (0-100 nM). Data are presented as mean ± SEM from n = 5 (BT-474 and SK-BR-3) and n = 6 (MCF7) independent experiments. C. SPRi analysis of the binding kinetics between HER2 and trastuzumab, pertuzumab, m66 and r40. Antibodies (0.5 mg/mL) were immobilized on a 3D optical crosslinking biosensor chip. HER2 was injected at concentrations ranging from 1 to 128 nM. Association and dissociation phases were monitored for 285 s and 930 s, respectively, using a PlexArray ® HT system.

Article Snippet: The human breast cancer cell lines BT-474, SK-BR-3 and MCF7 were obtained from the American Type Culture Collection (ATCC).

Techniques: Serial Dilution, CCK-8 Assay, Inhibition, Binding Assay, Injection

Journal: PLOS One

Article Title: Inhibition of HER2 signaling and breast cancer cell growth with a novel antibody targeting HER2 ECD III/IV

doi: 10.1371/journal.pone.0338127

Figure Lengend Snippet: Antibody binding to (A) BT-474, (B) SK-BR-3 and (C) MCF7 was evaluated by flow cytometry. Cells were incubated with 267 nM of each antibody and detected using fluorescence-labeled secondary antibodies.

Article Snippet: The human breast cancer cell lines BT-474, SK-BR-3 and MCF7 were obtained from the American Type Culture Collection (ATCC).

Techniques: Binding Assay, Flow Cytometry, Incubation, Fluorescence, Labeling

Journal: PLOS One

Article Title: Inhibition of HER2 signaling and breast cancer cell growth with a novel antibody targeting HER2 ECD III/IV

doi: 10.1371/journal.pone.0338127

Figure Lengend Snippet: A-C. Immunoblots showing HER2 downstream signaling in (A) MCF7, (B) BT-474 and (C) SK-BR-3 cells treated for 24 h with 10 μg/mL of the indicated antibodies. Whole-cell lysates were immunoblotted with antibodies against the indicated proteins. For combination treatments, the indicated concentration refers to each antibody individually. The numerical values shown in the figures represent normalized quantitative data.

Article Snippet: The human breast cancer cell lines BT-474, SK-BR-3 and MCF7 were obtained from the American Type Culture Collection (ATCC).

Techniques: Western Blot, Concentration Assay

Journal: PLOS One

Article Title: Inhibition of HER2 signaling and breast cancer cell growth with a novel antibody targeting HER2 ECD III/IV

doi: 10.1371/journal.pone.0338127

Figure Lengend Snippet: A-C. Immunoblots showing HER2 downstream signaling in (A) MCF7, (B) BT-474 and (C) SK-BR-3 treated with 10 μg/mL of the indicated antibodies for 24 h, followed by stimulation with 5 nM EGF for 10 min. D-F. Corresponding immunoblots of (D) MCF7, (E) BT-474 and (F) SK-BR-3 cells treated as above but stimulated with 0.5 nM NRG1 for 10 min. The numerical values shown in the figures represent normalized quantitative data.

Article Snippet: The human breast cancer cell lines BT-474, SK-BR-3 and MCF7 were obtained from the American Type Culture Collection (ATCC).

Techniques: Western Blot

Journal: Frontiers in Drug Delivery

Article Title: Anti-HER2-targeted therapies: effects on human in vitro blood-brain barrier models

doi: 10.3389/fddev.2025.1700455

Figure Lengend Snippet: Characterization of selected in vitro models and cell lines. The transendothelial electrical resistance (TEER) and permeability coefficient (Pe) for fluorescein were measured in the immortalized endothelial cell line hCMEC/D3 and in CD34+-derived brain-like endothelial cells (BLECs) (A) . Subsequently, the tight junctions were immunostained with an anti-Zonlula Occludens (ZO)-1 antibody. Top image: BLECs, bottom image: hCMEC/D3, scale bar 20 µm (B) . Messenger RNA expression levels of HER2 (encoded by the ERBB2 gene) (C) and epidermal growth factor receptor (EGFR) (D) in brain microvascular endothelial cell line hCMEC/D3 and BLECs, as well as in the breast cancer cell lines HCC1806 and BT 474 were measured by qPCR. Data are presented as means ± standard deviation (n = 3). ns, not significant, *p < 0.05, ***p < 0.001, ****p < 0.0001.

Article Snippet: HER-2 positive breast cancer cell line BT 474 (HTB-20, ATCC) was grown in MEM medium (Thermo Fisehr Scientific, 51200-038) supplemented with 10% fetal calf serum (FCS), L-glutamine and penicillin/streptomycin.

Techniques: In Vitro, Permeability, Derivative Assay, RNA Expression, Standard Deviation

Journal: Frontiers in Drug Delivery

Article Title: Anti-HER2-targeted therapies: effects on human in vitro blood-brain barrier models

doi: 10.3389/fddev.2025.1700455

Figure Lengend Snippet: Cell viability in breast cancer cell lines. The HER2-positive BT 474 cell line (A) and the triple-negative HC1806 breast cancer cell lines (B) were treated with 5, 50, 5,000 ng/mL trastuzumab, pertuzumab, trastuzumab/pertuzumab, lapatinib and tucatinib for 24 or 48 h followed by cell viability assay. Data are presented as means ± standard deviation and expressed as the fold of the untreated control, which was set at 1 (n = 3). *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001 compared to the untreated control.

Article Snippet: HER-2 positive breast cancer cell line BT 474 (HTB-20, ATCC) was grown in MEM medium (Thermo Fisehr Scientific, 51200-038) supplemented with 10% fetal calf serum (FCS), L-glutamine and penicillin/streptomycin.

Techniques: Viability Assay, Standard Deviation, Control

Journal: Science Advances

Article Title: FSP1 and histone deacetylases suppress cancer persister cell ferroptosis

doi: 10.1126/sciadv.aea8771

Figure Lengend Snippet: ( A ) UMAP of PC9 parental and persister cells treated with or without 1 μM RSL3 for 24 hours. ( B ) Enriched Hallmarks gene sets in persister cells treated with or without RSL3. Positive normalized enrichment score (NES) values indicate enrichment in RSL3-treated persister cells. TNFα, tumor necrosis factor–α; NF-κB, nuclear factor κB; IL-6, interleukin-6; JAK, Janus kinase; STAT3, signal transducer and activator of transcription 3; UV, ultraviolet. ( C ) UMAP of PC9 persister cells treated with and without RSL3. ( D ) Pseudotime analysis of PC9 persister cells treated with and without RSL3. Solid black line indicates the estimated trajectory across cell states. ( E ) UMAP of PC9 persister cells treated with and without RSL3 colored by cluster. ( F ) Ferroptosis driver gene set signature score across clusters in (E). ( G ) Ferroptosis suppressor gene set signature score across clusters in (E). [(F) and (G)] P values calculated with Mann-Whitney test. ( H ) PC9 parental, persister, and persister cells treated with 500 nM RSL3 for 24 hours were analyzed for oxygen consumption rate (OCR). A total of 1.25 μM oligomycin (Oligo), 1 μM carbonyl cyanide p -trifluoromethoxyphenylhydrazone (FCCP), and 1 μM rotenone plus 1 μM antimycin A (R + A). n = 3 biological replicates; mean ± SEM is shown; P values calculated between parental and persister cell conditions (stars) or between persister cells and RSL3-treated persister cells (crosses) using two-tailed Student’s t test. * P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001, †† P < 0.01, and ††† P < 0.001. ( I ) PC9 parental and persister cells derived from 2.5 μM erlotinib analyzed for mitochondrial ROS. Par, parental; pers, persister. ( J and K ) PC9 persister cells derived from 2.5 μM erlotinib (J) and A375 persister cells derived from 250 nM dabrafenib and 25 nM trametinib (K) in combination with mitoTEMPO were treated with 500 nM RSL3 for 24 hours. Viability was normalized to the respective mitoTEMPO-treated persister cells without RSL3 treatment. [(I) to (K)] n = 3 biological replicates; mean ± SD is shown; P values calculated with two-tailed Student’s t test.

Article Snippet: BT474 (HTB-20) and A375 (CRL-1619) cells were purchased from American Type Culture Collection.

Techniques: MANN-WHITNEY, Two Tailed Test, Derivative Assay

Journal: Science Advances

Article Title: FSP1 and histone deacetylases suppress cancer persister cell ferroptosis

doi: 10.1126/sciadv.aea8771

Figure Lengend Snippet: ( A ) Parental and persister cells analyzed for NRF2, KEAP1, and system x c − components SLC7A11 and SLC3A2 expression. ( B and C ) A375 and PC9 parental and persister cells analyzed for reduced GSH or total GSH (GSSG) levels. ( D ) Protein expression of ferroptosis suppressor genes in parental and persister cells. ( E ) FSP1 mRNA expression in PC9 parental and persister cells treated with and without RSL3. P values calculated with the Wilcoxon rank sum test with Bonferroni correction. ( F and G ) PC9 (F) and A375 (G) persister cells cotreated with 1 μM FSP1 inhibitor, iFSP1, 50 nM RSL3, or both for 24 hours. ( H and I ) PC9 (H) and A375 (I) viability of parental cells and persister cells after treatment with the combination of 1 μM iFSP1 with 50 nM RSL3 for 24 hours. [(B), (C), and (F) to (I)] n = 3 biological replicates; mean ± SD is shown; P values calculated with two-tailed Student’s t test. ns, not significant.

Article Snippet: BT474 (HTB-20) and A375 (CRL-1619) cells were purchased from American Type Culture Collection.

Techniques: Expressing, Two Tailed Test

Journal: Science Advances

Article Title: FSP1 and histone deacetylases suppress cancer persister cell ferroptosis

doi: 10.1126/sciadv.aea8771

Figure Lengend Snippet: ( A to F ) Synergy heatmaps between GPX4 inhibitor RSL3 and HDAC inhibitors panobinostat and vorinostat following 24-hour cotreatment. Bliss synergy score calculated with SynergyFinder 3.0. Red color and positive scores indicate synergy, and green color and negative scores indicate buffering. [(A) and (B)] PC9 parental cells and persister cells derived from 50 nM erlotinib. [(C) and (D)] A375 parental cells and persister cells derived from 10 nM dabrafenib with 1 nM trametinib. [(E) and (F)] BT474 parental cells and persister cells derived from 2 μM lapatinib. ( G to J ) Prederived PC9 or A375 persister cells were treated for 48 hours with a nontoxic concentration of HDAC inhibitor (see fig. S7), rinsed, and then treated with RSL3 for 24 hours while maintained under targeted therapy treatment. Data normalized to untreated persister cells. Concentration and HDAC inhibitor used were as follows: (G) 7.5 nM panobinostat, (H) 5 nM panobinostat, (I) 100 nM vorinostat, and (J) 1 μM vorinostat. RSL3 concentrations used were as follows: (G) 150 nM, (H) 100 nM, (I) 150 nM, and (J) 80 nM. n = 3 biological replicates; mean ± SD is shown; P values calculated with two-tailed Student’s t test.

Article Snippet: BT474 (HTB-20) and A375 (CRL-1619) cells were purchased from American Type Culture Collection.

Techniques: Derivative Assay, Concentration Assay, Two Tailed Test

Journal: Science Advances

Article Title: FSP1 and histone deacetylases suppress cancer persister cell ferroptosis

doi: 10.1126/sciadv.aea8771

Figure Lengend Snippet: ( A ) UMAP of PC9 parental and persister cells treated with or without HDAC inhibitor panobinostat for 48 hours. ( B ) Enriched Hallmarks gene sets between persister cells treated with and without panobinostat. Positive NES values indicate gene sets enriched in persister cells treated with panobinostat. ( C and D ) Treatment with panobinostat does not decrease GSH levels in PC9 (7.5 nM panobinostat) or A375 (5 nM panobinostat) persister cells. Buthionine sulfoximine (BSO; 1 mM) was used as a positive control for GSH depletion. ( E ) Ferroptosis sensitization of PC9 persister cells from treatment with panobinostat is not inhibited by GSH ethyl ester (GSHee, 1 mM). ( F ) PC9 persister cell treatment with panobinostat decreases rather than increases intracellular iron. ( G and H ) Panobinostat treatment of PC9 persister cells increases total cellular ROS (G) and mitochondrial ROS (H). ( I and J ) PC9 persister cells derived from 2.5 μM erlotinib (I) and A375 persister cells derived from 250 nM dabrafenib and 25 nM trametinib (J) were cotreated with mitoTEMPO and were treated with 7.5 and 5 nM panobinostat, respectively, for 48 hours with and without 500 nM RSL3 for 24 hours. Viability was normalized to the viability of targeted therapy with mitoTEMPO and panobinostat without RSL3. [(C) to (J)] n = 3 biological replicates; mean ± SD is shown; P values calculated with two-tailed Student’s t test.

Article Snippet: BT474 (HTB-20) and A375 (CRL-1619) cells were purchased from American Type Culture Collection.

Techniques: Positive Control, Derivative Assay, Two Tailed Test