triple negative breast cancer cell line hcc1806  (ATCC)

Journal: NeuroSci

Article Title: The Knockout of Protocadherin Gamma C3 (PCDHGC3) in Breast Cancer and Melanoma Cell Lines Leads to Increased Adhesion of Knockout Cells to Brain Microvascular Endothelial Cells

doi: 10.3390/neurosci7020047

Figure Lengend Snippet: Generation of PCDHGC3 knockout in breast cancer and melanoma cell lines. Western blot analysis of wild type (WT), control (C; transfected with the PCDH2 HDR Plasmid (h2) containing a puromycin resistance gene) and knockout (KO) HCC1806 breast cancer cell line ( a ) and A2058 melanoma cell line ( b ). β-Actin served as an endogenous control.

Article Snippet: Triple-negative breast cancer cell line HCC1806 (CRL-2335, ATCC, Manassas, VA, USA) was cultured in RPMI medium (R7509-500ML, Sigma-Aldrich, St. Louis, MO, USA) containing 10% FCS, L-glutamine and penicillin/streptomycin.

Techniques: Knock-Out, Western Blot, Control, Transfection, Plasmid Preparation

Journal: NeuroSci

Article Title: The Knockout of Protocadherin Gamma C3 (PCDHGC3) in Breast Cancer and Melanoma Cell Lines Leads to Increased Adhesion of Knockout Cells to Brain Microvascular Endothelial Cells

doi: 10.3390/neurosci7020047

Figure Lengend Snippet: Cell proliferation assay in control and PCDHGC3 knockout cells. Proliferation rate of control and PCDHGC3 knockout (KO) HCC1806 breast cancer cells ( a ) and of control and PCDHGC3 KO A2058 melanoma cells ( b ). **** = p < 0.0001, unpaired t test.

Article Snippet: Triple-negative breast cancer cell line HCC1806 (CRL-2335, ATCC, Manassas, VA, USA) was cultured in RPMI medium (R7509-500ML, Sigma-Aldrich, St. Louis, MO, USA) containing 10% FCS, L-glutamine and penicillin/streptomycin.

Techniques: Proliferation Assay, Control, Knock-Out

Journal: NeuroSci

Article Title: The Knockout of Protocadherin Gamma C3 (PCDHGC3) in Breast Cancer and Melanoma Cell Lines Leads to Increased Adhesion of Knockout Cells to Brain Microvascular Endothelial Cells

doi: 10.3390/neurosci7020047

Figure Lengend Snippet: Relative adhesion of PCDHGC3 knockout breast cancer and melanoma cells to human in vitro BBB models. Adhesion measurements of HCC1806 PCDHGC3 knockout (KO) and control cells to hCMEC/D3 ( a ) and BLECs ( b ) after 30, 60, and 120 min. Adhesion measurements of A2058 PCDHGC3 knockout (KO) and control cells to hCMEC/D3 ( c ) and BLECs ( d ) after 30, 60, and 120 min. Control cell adhesion was measured at each time point; however, for clarity, only the measurement after 30 min is shown. Data are presented as mean relative adhesion versus control with standard deviation, **** = p ≤ 0.0001, one-way ANOVA test.

Article Snippet: Triple-negative breast cancer cell line HCC1806 (CRL-2335, ATCC, Manassas, VA, USA) was cultured in RPMI medium (R7509-500ML, Sigma-Aldrich, St. Louis, MO, USA) containing 10% FCS, L-glutamine and penicillin/streptomycin.

Techniques: Knock-Out, In Vitro, Control, Standard Deviation

Journal: NeuroSci

Article Title: The Knockout of Protocadherin Gamma C3 (PCDHGC3) in Breast Cancer and Melanoma Cell Lines Leads to Increased Adhesion of Knockout Cells to Brain Microvascular Endothelial Cells

doi: 10.3390/neurosci7020047

Figure Lengend Snippet: PCDHGC3 KO leads to stronger invasion of PCDHGC3 knockout breast cancer and melanoma cells. HCC1806 PCDHGC3 knockout (KO) and control cells ( a ) and A2058 PCDHGC3 knockout (KO) and control ( b ) invaded for 48 h through Transwells coated with Matrigel. The number of invaded cells is shown. Data are presented as mean cell number with standard deviation, * = p ≤ 0.05, unpaired t -test.

Article Snippet: Triple-negative breast cancer cell line HCC1806 (CRL-2335, ATCC, Manassas, VA, USA) was cultured in RPMI medium (R7509-500ML, Sigma-Aldrich, St. Louis, MO, USA) containing 10% FCS, L-glutamine and penicillin/streptomycin.

Techniques: Knock-Out, Control, Standard Deviation

Journal: NeuroSci

Article Title: The Knockout of Protocadherin Gamma C3 (PCDHGC3) in Breast Cancer and Melanoma Cell Lines Leads to Increased Adhesion of Knockout Cells to Brain Microvascular Endothelial Cells

doi: 10.3390/neurosci7020047

Figure Lengend Snippet: Relative expression of target genes in PCDHGC3 KO breast cancer and melanoma cells. The relative expression (RQ value) of each target in PCDHGC3 KO HCC1806 ( a ) and PCDHGC3 KO A2058 ( b ) cells relative to control cells is shown. A RQ value < 1.0 indicates decreased expression, a RQ value > 1.0 indicates increased expression compared to the control cells. The means with standard deviation are shown as the fold of the control. * = p ≤ 0.05, ** = p ≤ 0.01, *** = p ≤ 0.001, **** = p ≤ 0.0001, unpaired t -test.

Article Snippet: Triple-negative breast cancer cell line HCC1806 (CRL-2335, ATCC, Manassas, VA, USA) was cultured in RPMI medium (R7509-500ML, Sigma-Aldrich, St. Louis, MO, USA) containing 10% FCS, L-glutamine and penicillin/streptomycin.

Techniques: Expressing, Control, Standard Deviation

Journal: NeuroSci

Article Title: The Knockout of Protocadherin Gamma C3 (PCDHGC3) in Breast Cancer and Melanoma Cell Lines Leads to Increased Adhesion of Knockout Cells to Brain Microvascular Endothelial Cells

doi: 10.3390/neurosci7020047

Figure Lengend Snippet: Matrix metalloproteinase (MMP) activity in cell culture medium of PCDHGC3 knockout (KO) breast cancer and melanoma cells. Fluorescence signal of MMP-substrate (SB) cleavage in the cell culture medium of PCDHGC3 KO HCC1806 ( a ) and PCDHGC3 KO A2058 ( b ) cells and control cells is expressed as relative fluorescence units (RFU) ± standard deviation. ** = p < 0.01, *** = p < 0.001, unpaired t test.

Article Snippet: Triple-negative breast cancer cell line HCC1806 (CRL-2335, ATCC, Manassas, VA, USA) was cultured in RPMI medium (R7509-500ML, Sigma-Aldrich, St. Louis, MO, USA) containing 10% FCS, L-glutamine and penicillin/streptomycin.

Techniques: Activity Assay, Cell Culture, Knock-Out, Fluorescence, Control, Standard Deviation

Journal: Cancers

Article Title: Hypoxia-Inducible Factor-1α, a Novel Molecular Target for a 2-Aminopyrrole Derivative: Biological and Molecular Modeling Study

doi: 10.3390/cancers18010115

Figure Lengend Snippet: Anti-proliferative effects of 2-ANPC in epithelial cancer cell lines. Changes in growth kinetics of HCC1806, MDA-MB-231, H1299, and PC-3 cells treated with 2-ANPC, PTX (positive control) and solvent DMSO (negative control). Cells (0.5 × 10 5 /mL) were seeded into the wells of an E-Plate L8 PET cassette and installed in the iCELLigence cell growth kinetics system (ACEA Biosciences, San Diego, CA, USA). Cells were allowed to attach and grow for the following 24 h. Afterwards, 2-ANPC, PTX, or DMSO was introduced into the cell culture. Cell proliferation index values were recorded every hour throughout the experiment. RTCA Software version 1.0 (ACEA Biosciences, Inc., San Diego, CA, USA) was used to analyze the data.

Article Snippet: Triple negative breast cancer HCC1806, MDA-MB-231, and 4T1; non-small cell lung carcinoma H1299; and prostate cancer PC-3 cell lines were obtained from the American Type Culture Collection (ATCC, Manassas, VA, USA).

Techniques: Positive Control, Solvent, Negative Control, Cell Culture, Software

Journal: Cancers

Article Title: Hypoxia-Inducible Factor-1α, a Novel Molecular Target for a 2-Aminopyrrole Derivative: Biological and Molecular Modeling Study

doi: 10.3390/cancers18010115

Figure Lengend Snippet: Pro-apoptotic activity of 2-ANPC in epithelial cancer cell lines. HCC1806, MDA-MB-231, H1299, and PC-3 cells were treated with 2-ANPC (10 µM) for 48 h and subjected to Western blot analysis to examine the expression of apoptotic markers, including cleaved PARP and cleaved caspase-3. Actin staining was used to show the comparable amounts of protein loaded into each sample. Representative Western blotting results from 3 independent experiments are shown. The figure on the right shows the quantification of HIF-1α expression in cells based on the mean pixel density. The uncropped original Western blotting images can be found in .Values are means ± SD, n = 3. (*— p < 0.01; one-way ANOVA).

Article Snippet: Triple negative breast cancer HCC1806, MDA-MB-231, and 4T1; non-small cell lung carcinoma H1299; and prostate cancer PC-3 cell lines were obtained from the American Type Culture Collection (ATCC, Manassas, VA, USA).

Techniques: Activity Assay, Western Blot, Expressing, Staining

Journal: Cancers

Article Title: Hypoxia-Inducible Factor-1α, a Novel Molecular Target for a 2-Aminopyrrole Derivative: Biological and Molecular Modeling Study

doi: 10.3390/cancers18010115

Figure Lengend Snippet: 2-ANPC decreases expression of HIF-1α in epithelial cancer cell lines by reducing its stability and promoting the proteasome-dependent degradation. ( A ) HCC1806, MDA-MB-231, H1299, and PC-3 cells were treated with 2-ANPC (10 µM) for 48 h and subjected to Western blotting analysis three times to examine the expression of HIF-1α. Actin staining was used to show comparable amounts of protein loaded into each sample. The figure on the right shows the quantification of HIF-1α expression in cells based on the mean pixel density. Values are means ± SD; n = 3. An asterisk indicates a significant difference compared to relevant controls ( p < 0.05; one-way ANOVA). ( B ) Changes in the relative expression level of HIF-1α mRNA in epithelial cancer cells (HCC1806, MDA-MB-231, H1299, PC-3) treated with 2-ANPC (10 µM—48 h), as determined by quantitative RT-PCR. As an internal control, glyceraldehyde-3-phosphate dehydrogenase ( GAPDH ) amplification was used. Data are presented as median ± standard deviation (SD). Significant differences at p < 0.05 (*), p < 0.001 (**) from n ≥ 3 using an unpaired Student’s t -test are presented. ns – non significant ( C ) To examine the impact of 2-ANPC on proteasome degradation of HIF-1α, PC-3 prostate cancer cells were treated with 2-ANPC in the presence of MG-132 (2.5 μM) for 6 h and were subjected to immunoblotting for HIF-1α and actin as a loading control. Representative Western blotting results from 3 independent experiments are shown. The figure on the right shows the quantification of HIF-1α expression in cells based on the mean pixel density. Values are means ± SD; n = 3. An asterisk indicates significant difference compared to relevant controls (*— p < 0.01; one-way ANOVA).

Article Snippet: Triple negative breast cancer HCC1806, MDA-MB-231, and 4T1; non-small cell lung carcinoma H1299; and prostate cancer PC-3 cell lines were obtained from the American Type Culture Collection (ATCC, Manassas, VA, USA).

Techniques: Expressing, Western Blot, Staining, Quantitative RT-PCR, Control, Amplification, Standard Deviation

Journal: Cancers

Article Title: Hypoxia-Inducible Factor-1α, a Novel Molecular Target for a 2-Aminopyrrole Derivative: Biological and Molecular Modeling Study

doi: 10.3390/cancers18010115

Figure Lengend Snippet: 2-ANPC decreases HIF-1α’s expression under hypoxic conditions. Effects of CoCl 2 treatment on HIF-1α expression on protein ( A , B ) and mRNA ( C ) levels. Cancer cells (HCC1806, MDA-MB-231, PC-3) were incubated in the absence (control) or in the presence of 100 μM CoCl 2 for 24 h. 2-ANPC (10 μM) was introduced into the cell cultures simultaneously with CoCl 2 . HIF-1α protein expression was measured by Western blotting ( A ), and HIF-1α mRNA expression was determined by RT-PCR ( C ). ( B ) Quantification of HIF-1α expression in cells based on the average pixel density. Values are presented as mean ± standard deviation; n = 3. Statistically significant differences between the respective groups are marked with an asterisk (*— p < 0.05; **— p < 0.01; one-way ANOVA).

Article Snippet: Triple negative breast cancer HCC1806, MDA-MB-231, and 4T1; non-small cell lung carcinoma H1299; and prostate cancer PC-3 cell lines were obtained from the American Type Culture Collection (ATCC, Manassas, VA, USA).

Techniques: Expressing, Incubation, Control, Western Blot, Reverse Transcription Polymerase Chain Reaction, Standard Deviation

Journal: Cancers

Article Title: Anticancer Activity of Encapsulated Pearl Millet Polyphenol-Rich Extract against Proliferating and Non-Proliferating Breast Cancer Cells In Vitro

doi: 10.3390/cancers16091750

Figure Lengend Snippet: The effects of polyphenolic fractions dissolved in DMSO (PD, ( top )), ethanol (PE, ( middle )), and lipids (PL, ( bottom )) extracted from pearl millet grains on metabolic activity of three breast cancer cell lines, namely triple negative HCC1806, ER-positive HCC1428, and HER2-positive AU565 cells. Cells were treated with pearl millet extracts at the concentrations of 10, 20, 40, and 60 μg/mL for 24 h. Metabolic activity was assayed using MTT test. Untreated control (CTR) is considered as 100%. The effects of solvents (DMSO, EtOH) were also tested. Bars indicate SD, n = 6, *** p < 0.001, ** p < 0.01, * p < 0.05 compared to CTR (ANOVA and Dunnett’s a posteriori test).

Article Snippet: The following breast cancer cell lines, namely triple negative breast cancer (TNBC) HCC1806 (CRL-2335 TM ), ER-positive HCC1428 (CRL-2327 TM ), and HER2-positive AU565 (CRL-2351 TM ) (ATCC, Manassas, VA, USA), were grown in RPMI-1640 medium (Corning, Tewksbury, MA, USA) supplemented with 10% FBS and antibiotics (100 U/mL penicillin, 0.1 mg/mL streptomycin, and 0.25 g/mL amphotericin).

Techniques: Activity Assay, Control

Journal: Cancers

Article Title: Anticancer Activity of Encapsulated Pearl Millet Polyphenol-Rich Extract against Proliferating and Non-Proliferating Breast Cancer Cells In Vitro

doi: 10.3390/cancers16091750

Figure Lengend Snippet: The effects of encapsulated polyphenolic extracts (PDMC and PEMC) on metabolic activity of three breast cancer cell lines, namely HCC1806, HCC1428, and AU565, and corresponding non-tumorigenic MCF10F cells. Metabolic activity was analyzed using MTT test. Cells were treated with pearl millet-encapsulated polyphenolic extracts at the concentrations of 0.62, 1.25, and 2.5% for 24 h. The effects of microcapsule-based drug delivery system (empty MC) were also tested. The metabolic activity of untreated control (CTR) is considered as 100%. Bars indicate SD, n = 6, *** p < 0.001 compared to CTR (ANOVA and Dunnett’s a posteriori test), ### p < 0.001, ## p < 0.01 compared to MC treatment (ANOVA and Tukey’s a posteriori test). CTR, control conditions; MC, treatment with empty microcapsules; PDMC, treatment with encapsulated pearl millet polyphenolic extract dissolved in DMSO; PEMC, treatment with encapsulated pearl millet polyphenolic extract dissolved in EtOH.

Article Snippet: The following breast cancer cell lines, namely triple negative breast cancer (TNBC) HCC1806 (CRL-2335 TM ), ER-positive HCC1428 (CRL-2327 TM ), and HER2-positive AU565 (CRL-2351 TM ) (ATCC, Manassas, VA, USA), were grown in RPMI-1640 medium (Corning, Tewksbury, MA, USA) supplemented with 10% FBS and antibiotics (100 U/mL penicillin, 0.1 mg/mL streptomycin, and 0.25 g/mL amphotericin).

Techniques: Activity Assay, Control

Journal: Cancers

Article Title: Anticancer Activity of Encapsulated Pearl Millet Polyphenol-Rich Extract against Proliferating and Non-Proliferating Breast Cancer Cells In Vitro

doi: 10.3390/cancers16091750

Figure Lengend Snippet: PDMC- and PEMC-mediated apoptotic cell death in three breast cancer cell lines with different receptor status, namely HCC1806, HCC1428, and AU565 ( B ) in comparison to PDMC- and PEMC-associated effects in non-tumorigenic MCF10F cells ( A ). Cells were treated with encapsulated pearl millet polyphenolic extracts at the concentrations of 0.62 and 2.5% (normal cells, ( A )) and 2.5% (cancer cells, ( B )) for 24 h. Apoptosis was assessed as phosphatidylserine externalization using Annexin V staining and flow cytometry. Bars indicate SD, n = 3, *** p < 0.001, ** p < 0.01, * p < 0.05 compared to CTR (ANOVA and Dunnett’s a posteriori test), ### p < 0.001 compared to MC treatment (ANOVA and Tukey’s a posteriori test). Representative dot-plots are also shown. CTR, control conditions; MC, treatment with empty microcapsules; PDMC, treatment with encapsulated pearl millet polyphenolic extract dissolved in DMSO; PEMC, treatment with encapsulated pearl millet polyphenolic extract dissolved in EtOH.

Article Snippet: The following breast cancer cell lines, namely triple negative breast cancer (TNBC) HCC1806 (CRL-2335 TM ), ER-positive HCC1428 (CRL-2327 TM ), and HER2-positive AU565 (CRL-2351 TM ) (ATCC, Manassas, VA, USA), were grown in RPMI-1640 medium (Corning, Tewksbury, MA, USA) supplemented with 10% FBS and antibiotics (100 U/mL penicillin, 0.1 mg/mL streptomycin, and 0.25 g/mL amphotericin).

Techniques: Comparison, Staining, Flow Cytometry, Control

Journal: Cancers

Article Title: Anticancer Activity of Encapsulated Pearl Millet Polyphenol-Rich Extract against Proliferating and Non-Proliferating Breast Cancer Cells In Vitro

doi: 10.3390/cancers16091750

Figure Lengend Snippet: The uptake of microcapsules ( A ) and encapsulated polyphenolic extract-mediated anticancer effects in doxorubicin-induced senescent breast cancer cells HCC1806, HCC1428, and AU565 ( B , C ). Senescent breast cancer cells were treated with 0.62% microcapsules for 24 h. ( A ) The uptake of microcapsules was studied using fluorescein-containing microcapsules (MC*, PDMC*, and PEMC*) and imaging cytometry. The results are presented as relative fluorescence units (RFU). Box and whisker plots are shown, n = 6, *** p < 0.001 compared to CTR (ANOVA and Dunnett’s a posteriori test). ( B ) The analysis of phosphatidylserine externalization, a marker of apoptosis using Annexin V staining and flow cytometry. Bars indicate SD, n = 3, *** p < 0.001, ** p < 0.01 compared to CTR (ANOVA and Dunnett’s a posteriori test). Representative dot-plots are also presented. ( C , D ) The analysis of the levels of cytoplasmic caspase 9 (Casp9) and caspase 3 (Casp3) (apoptotic markers), nuclear p21 (cell cycle inhibitor), and cytoplasmic pro-inflammatory cytokine IL8 using dedicated antibodies and imaging cytometry. ( C ) Data are presented as relative fluorescence units (RFU). Box and whisker plots are shown, n = 6, *** p < 0.001, ** p < 0.01, * p < 0.05 compared to CTR (ANOVA and Dunnett’s a posteriori test). ( D ) Representative microphotographs of non-treated (CTR) and treated AU565 cells (response) are also shown. Casp9, Casp3, p21, and IL8 immunosignals are presented in red (immunostaining with dedicated primary antibodies and fluorochrome conjugated secondary antibodies). Nuclei are presented in blue (Hoechst 33342 staining). Objective 20×. CTR, control conditions; MC, treatment with empty microcapsules; PDMC, treatment with encapsulated pearl millet polyphenolic extract dissolved in DMSO; PEMC, treatment with encapsulated pearl millet polyphenolic extract dissolved in EtOH.

Article Snippet: The following breast cancer cell lines, namely triple negative breast cancer (TNBC) HCC1806 (CRL-2335 TM ), ER-positive HCC1428 (CRL-2327 TM ), and HER2-positive AU565 (CRL-2351 TM ) (ATCC, Manassas, VA, USA), were grown in RPMI-1640 medium (Corning, Tewksbury, MA, USA) supplemented with 10% FBS and antibiotics (100 U/mL penicillin, 0.1 mg/mL streptomycin, and 0.25 g/mL amphotericin).

Techniques: Imaging, Cytometry, Fluorescence, Whisker Assay, Marker, Staining, Flow Cytometry, Immunostaining, Control

Journal: Cancers

Article Title: Anticancer Activity of Encapsulated Pearl Millet Polyphenol-Rich Extract against Proliferating and Non-Proliferating Breast Cancer Cells In Vitro

doi: 10.3390/cancers16091750

Figure Lengend Snippet: PDMC- and PEMC-mediated changes in the levels of autophagic markers beclin-1 (BECN1) and LC3b in doxorubicin-induced senescent breast cancer cells HCC1806, HCC1428, and AU565. ( A , B ) Senescent breast cancer cells were treated with 0.62% microcapsules for 24 h. The levels of cytoplasmic BECN1 and LC3b were analyzed using dedicated antibodies and imaging cytometry. ( A ) Data are presented as relative fluorescence units (RFU). Box and whisker plots are presented, n = 6, *** p < 0.001, * p < 0.05 compared to CTR (ANOVA and Dunnett’s a posteriori test). ( B ) Representative microphotographs of non-treated (CTR) and treated AU565 cells (response) are also shown. BECN1 and LC3b immunosignals are presented in green and red, respectively (immunostaining with dedicated primary antibodies and fluorochrome conjugated secondary antibodies). Nuclei are presented in blue (Hoechst 33342 staining). Objective 20×. CTR, control conditions; MC, treatment with empty microcapsules; PDMC, treatment with encapsulated pearl millet polyphenolic extract dissolved in DMSO; PEMC, treatment with encapsulated pearl millet polyphenolic extract dissolved in EtOH.

Article Snippet: The following breast cancer cell lines, namely triple negative breast cancer (TNBC) HCC1806 (CRL-2335 TM ), ER-positive HCC1428 (CRL-2327 TM ), and HER2-positive AU565 (CRL-2351 TM ) (ATCC, Manassas, VA, USA), were grown in RPMI-1640 medium (Corning, Tewksbury, MA, USA) supplemented with 10% FBS and antibiotics (100 U/mL penicillin, 0.1 mg/mL streptomycin, and 0.25 g/mL amphotericin).

Techniques: Imaging, Cytometry, Fluorescence, Whisker Assay, Immunostaining, Staining, Control

Journal: Cancers

Article Title: Anticancer Activity of Encapsulated Pearl Millet Polyphenol-Rich Extract against Proliferating and Non-Proliferating Breast Cancer Cells In Vitro

doi: 10.3390/cancers16091750

Figure Lengend Snippet: Diverse cell death responses in drug-induced senescent breast cancer cells after treatment with encapsulated pearl millet polyphenolic extracts (PDMC and PEMC). Three phenotypically different cellular models of breast cancer were considered, namely TNBC (HCC1806 cells), ER-positive (HCC1428 cells), and HER2-positive (AU565 cells). Encapsulated polyphenolic extract induced apoptosis in all three breast cancer cell lines. Necrosis was also observed in encapsulated polyphenolic extract-treated TNBC and HER2-positive cells, which was accompanied by increased levels of p21 and IL8. Encapsulated polyphenolic extract-induced decrease in cell viability was also accompanied by the activation of autophagic pathway, here cytotoxic autophagy, in HER2-positive cells. Encapsulated polyphenolic extract also promoted apoptotic cell death in proliferating breast cancer cells and pro-apoptotic activity of encapsulated polyphenolic extract was potentiated compared to the cytotoxic effects induced by free polyphenolic fractions. PDMC, treatment with encapsulated pearl millet polyphenolic extract dissolved in DMSO; PEMC, treatment with encapsulated pearl millet polyphenolic extract dissolved in EtOH.

Article Snippet: The following breast cancer cell lines, namely triple negative breast cancer (TNBC) HCC1806 (CRL-2335 TM ), ER-positive HCC1428 (CRL-2327 TM ), and HER2-positive AU565 (CRL-2351 TM ) (ATCC, Manassas, VA, USA), were grown in RPMI-1640 medium (Corning, Tewksbury, MA, USA) supplemented with 10% FBS and antibiotics (100 U/mL penicillin, 0.1 mg/mL streptomycin, and 0.25 g/mL amphotericin).

Techniques: Activation Assay, Activity Assay