mouse anti-her3  (Novocastra)

Journal: Cancers

Article Title: The Co-Expression and Cellular Location of HER Family Members, EGFRvIII, Putative Cancer Stem Cell Biomarkers CD44 and CD109 in Patients with Glioblastoma, and Their Impacts on Prognosis

doi: 10.3390/cancers17071221

Figure Lengend Snippet: Studies investigating the expression level and prognostic significance of HER2, HER3 and HER4 in patients with glioblastoma immunohistochemistry.

Article Snippet: (Nabika et al., 2010) [ ] , Mouse anti-HER3 (Novocastra), and HER4 (Lab vision) antibodies/scored by counting the numbering of positive cells per 1000 tumour cells and producing labelling index (i.e., < 30% as negative, >30% as positive). , 59 High grade astrocytoma HER3 (5.1%) HER4 (75%) , High expression of HER4 was associated with a poor prognosis ( p = 0.004) but no association between HER3 and prognosis.

Techniques: Expressing, Immunohistochemistry, Amplification, Over Expression, Staining, Marker, Biomarker Discovery

Journal: Breast Cancer Research : BCR

Article Title: A prospective study of HER3 expression pre and post neoadjuvant therapy of different breast cancer subtypes: implications for HER3 imaging therapy guidance

doi: 10.1186/s13058-024-01859-w

Figure Lengend Snippet: Survival analysis in the context of HER3 gene alteration. A Kaplan–Meier plots for disease-specific, progression free and overall survival of breast cancer showing HER3 gene alterations from 221 patients across 16 studies, using cBioPortal. B Mutation frequency in percentage of genes associated with HER3 gene signatures in breast cancer C alterations in EGFR, HER2, HER3 and ESR1 gene expression tendencies from patients with breast cancer D schematic representation of pathway signalling in patients with HER3 gene signature. Data was generated in ( http://cbioportal.org )

Article Snippet: Primary antibody incubation was performed overnight at 4 °C with the addition of rabbit anti-mouse HER3 antibody (12,780, Cell Signaling).

Techniques: Mutagenesis, Gene Expression, Generated

Journal: Breast Cancer Research : BCR

Article Title: A prospective study of HER3 expression pre and post neoadjuvant therapy of different breast cancer subtypes: implications for HER3 imaging therapy guidance

doi: 10.1186/s13058-024-01859-w

Figure Lengend Snippet: HER3 is variably expressed across different breast cancer subtypes at diagnosis. A Relative protein expression of HER3 in patients (n = 34) diagnosed with breast tumors and classified by hormone receptor and HER2 expression subtype with B mean HER3 expression based on receptor status and C patient frequency of low and high HER3 expressing tumors D Distribution of patients based on specific hormone status E Representative immunoblots of HER3 family receptors and downstream signaling demonstrating variable HER3 involvement in these breast cancer patients F Pearson correlation analysis showing a positive correlation between HER3 expression and phosphorylation of AKT in breast cancer patients G Pearson correlation between HER3 and phosphorylation of AKT as classified by intrinsic breast cancer subtype and H representative immunohistofluorescence analysis of HER3 expressing and non HER3 expressing breast cancer biopsies

Article Snippet: Primary antibody incubation was performed overnight at 4 °C with the addition of rabbit anti-mouse HER3 antibody (12,780, Cell Signaling).

Techniques: Biomarker Discovery, Expressing, Western Blot, Phospho-proteomics, Immunohistofluorescence

Journal: Breast Cancer Research : BCR

Article Title: A prospective study of HER3 expression pre and post neoadjuvant therapy of different breast cancer subtypes: implications for HER3 imaging therapy guidance

doi: 10.1186/s13058-024-01859-w

Figure Lengend Snippet: HER3 increases after neoadjuvant therapy in some but not all breast cancer tumors. A Patient characteristics and neoadjuvant regimens given B Representative immunoblots of HER3 and downstream signaling in patients pre and post neoadjuvant therapy and C Quantitative analysis of changes in these protein targets demonstrating variable response in the HER3 expression (n = 3). Data was analyzed using ANOVA analysis D Immunohistofluorescence of HER3 in patients receiving neoadjuvant therapy and E corresponding quantification analyzed using T-tests. * P < 0.05; ** P < 0.01, *** P < 0.001, ***** P < 0.0001

Article Snippet: Primary antibody incubation was performed overnight at 4 °C with the addition of rabbit anti-mouse HER3 antibody (12,780, Cell Signaling).

Techniques: Western Blot, Expressing, Immunohistofluorescence

Journal: Breast Cancer Research : BCR

Article Title: A prospective study of HER3 expression pre and post neoadjuvant therapy of different breast cancer subtypes: implications for HER3 imaging therapy guidance

doi: 10.1186/s13058-024-01859-w

Figure Lengend Snippet: Assessment of HER3 in patients from the GSE122630 study pre and post neoadjuvant therapy. A HER3 expression at baseline (T1), at two weeks during neoadjuvant therapy (T2), mid-neoadjuvant therapy (T3) and at surgical resection (T4) B HER3 expression per patient at all timepoints where a biopsy was available with insets showing an example of a patient with HER3 decrease and a patient with HER3 increase C Microenvironmental cell populations in patients with HER3 decrease and patients with HER3 increase at T4 (surgical resection)

Article Snippet: Primary antibody incubation was performed overnight at 4 °C with the addition of rabbit anti-mouse HER3 antibody (12,780, Cell Signaling).

Techniques: Expressing

Journal: Breast Cancer Research : BCR

Article Title: A prospective study of HER3 expression pre and post neoadjuvant therapy of different breast cancer subtypes: implications for HER3 imaging therapy guidance

doi: 10.1186/s13058-024-01859-w

Figure Lengend Snippet: Differential biologies in patients with HER3 increase post neoadjuvant therapy. A Differential gene expression in patients with HER3 decrease or HER3 increase at T4 (surgical resection). B Top 20 most upregulated and downregulated genesets in patients that showed a HER3 increase or decrease post neoadjuvant therapy. C Enrichment plots showing upregulation and downregulation of immune response in patients with HER3 decrease or HER3 increase post neoadjuvant therapy. D String network formed by the top 11 most upregulated genes in patients that showed a HER3 decrease post-neoadjuvant therapy

Article Snippet: Primary antibody incubation was performed overnight at 4 °C with the addition of rabbit anti-mouse HER3 antibody (12,780, Cell Signaling).

Techniques: Gene Expression

Journal: Breast Cancer Research : BCR

Article Title: A prospective study of HER3 expression pre and post neoadjuvant therapy of different breast cancer subtypes: implications for HER3 imaging therapy guidance

doi: 10.1186/s13058-024-01859-w

Figure Lengend Snippet: A Heatmap of the hallmark gene sets between patients with decreased or increased HER3 expression at T1 and T4 (n = 25) with red inset depicting the statistically significant hallmarks and B Boxplots showing comparison between patients with an increase or a decrease in HER3 post NAT in a select number of significantly differential pathways from the Hallmarks gene sets as analyzed using Kruskal–Wallis test. T -test between groups were carried out and P -values < 0.05 were considered statistically significant

Article Snippet: Primary antibody incubation was performed overnight at 4 °C with the addition of rabbit anti-mouse HER3 antibody (12,780, Cell Signaling).

Techniques: Expressing, Comparison

Journal: World Journal of Oncology

Article Title: Synergistic Effects of Neratinib in Combination With Palbociclib or Miransertib in Brain Cancer Cells

doi: 10.14740/wjon1873

Figure Lengend Snippet: Surface Expression of Various Growth Factor Receptors in Human Brain Cancer Cell Line (Determined by Flow Cytometry)

Article Snippet: The mouse monoclonal antibodies against HER3 (MAB3481), HER4 (MAB11311), ALK7 (MAB77491) and HGF R/c-MET (MAB3582) were purchased from R&D Systems (Oxford, UK).

Techniques: Expressing, Cytometry, Control, Fluorescence

Journal: Cancer Cell International

Article Title: HER3 functions as an effective therapeutic target in triple negative breast cancer to potentiate the antitumor activity of gefitinib and paclitaxel

doi: 10.1186/s12935-023-03055-w

Figure Lengend Snippet: Elevated expression of HER3 is frequently observed in TNBC clinical samples and cell lines and associates with poor clinical outcomes in TNBC patients. A , IHC assays were performed to examine HER3 protein expression in a tissue microarray (TMA) of TNBC specimens (n = 125). Representative images of negative (-), low (+), medium (++), and high (+++) staining intensity of HER3 expression were shown. The case number in each group was shown underneath. B , The heatmap showed HER3 mRNA expression in CCLE breast cancer cell lines ( http://software.broadinstitute.org/morpheus ). Red colors indicated TNBC cell lines. C , Analyses of overall survival (OS) or relapse-free survival (RFS) of basal-like breast cancer patients were performed using Kaplan-Meier Plotter ( https://kmplot.com/analysis/ ). The expression levels of HER3 mRNA were determined in TCGA-curated basal-like breast cancer datasets and used for log rank tests to compare the survival curves of those with high (red) or low (black) HER3 expression

Article Snippet: Antibodies used for flow cytometry analysis were mouse anti-HER3 mAb (10,201-MM01) from Sino Biological, Inc. (Beijing, China).

Techniques: Expressing, Microarray, Staining, Software

Journal: Cancer Cell International

Article Title: HER3 functions as an effective therapeutic target in triple negative breast cancer to potentiate the antitumor activity of gefitinib and paclitaxel

doi: 10.1186/s12935-023-03055-w

Figure Lengend Snippet: Specific knockdown of HER3 expression markedly inhibits TNBC cell proliferation and mammosphere formation in vitro and profoundly suppresses TNBC tumor growth in vivo. A and B , HCC1806, MDA-MB-468 (MDA-468), and MDA-MB-231 (MDA-231) cells were infected with lentivirus containing either control shRNA (shControl) or specific shRNA against HER3 (sh HER3 ) for 24 h. The cells were then seeded in 96-well plates, incubated at a regular cell culture incubator, and monitored for growth via an IncuCyte system for 64 h. The cell growth curves were generated by GraphPad Prism9 software with the data obtained through IncuCyte scanning every 4 h ( A ). The cells were then subjected to mammosphere formation assays. After 14 days, representative images of the mammospheres were shown and the number and size of those mammospheres were examined, quantified, and presented in the bar graphs ( B ). Data shows a representative of three independent experiments. Bars, SD. *, p < 0.05, **, p < 0.01, ***, p < 0.005. C , Luciferase-labelled HCC1806 cells were infected with lentivirus containing either control shRNA (shControl) or HER3 shRNA (sh HER3 ) for 24 h. The HCC1806-shControl or HCC1806-sh HER3 cells (5 × 10 5 ) were orthotopically inoculated into the left or right mammary fat pads of female nude mice, respectively (n = 4). In vivo bioluminescent imaging of the mammary tumors was obtained with the IVIS spectrum at day 1, 7, or 14 after cell inoculation. The graph showed the luciferase signal intensity of the mammary tumors obtained from the mice injected with HCC1806-shControl or HCC1806-sh HER3 cells at day 1 and 14. A significant difference was observed using a two-sided Student’s t-test

Article Snippet: Antibodies used for flow cytometry analysis were mouse anti-HER3 mAb (10,201-MM01) from Sino Biological, Inc. (Beijing, China).

Techniques: Expressing, In Vitro, In Vivo, Infection, shRNA, Incubation, Cell Culture, Generated, Software, Luciferase, Imaging, Injection

Journal: Cancer Cell International

Article Title: HER3 functions as an effective therapeutic target in triple negative breast cancer to potentiate the antitumor activity of gefitinib and paclitaxel

doi: 10.1186/s12935-023-03055-w

Figure Lengend Snippet: Our newly developed anti-HER3 mAb 4A7 recognizes cell membrane HER3, and exhibits profound activity to block HRGβ1-, but not SDF-1-induced TNBC cell migration. A , Our anti-HER3 mAb 4A7 was produced and purified as described in the materials and methods. 4A7 binding to membrane HER3 on SKBR3.B3.1 cells was examined by flow cytometry analysis. Staining with 2nd Ab only was used as a negative control. 10,201-MM01, a commercially available anti-HER3 mAb was used a positive control. Quantification of the positive staining with either 10,201-MM01 or 4A7 from three independent experiments was shown in the bar graphs. B , Equal amount of total protein extracts of HCC1806 cells were either examined by western blot analyses of HER3 and IGF-1R (Input) or subjected to IP (IP: Ab) using a control IgG or 4A7 and followed by western blot analyses of HER3 and IGF-1R. C-D , The specificity of 4A7 against HER3 was confirmed by its capability to block HRGβ1-induced promotion of TNBC cell migration. HCC1806 cells seeded in 96-well plates were pre-incubated with mitomycin C (2 µg/ml) prior to the scratch assays. The cells were then untreated or treated with HRGβ1 (50ng/ml) or HRGβ1 plus 4A7 (20ug/ml) and monitored by an IncuCyte system for cell migration ( C ). The same migration assays were performed with MDA-MB-231 cells in the presence of HRGβ1 (50ng/ml) or SDF-1 (200ng/ml) or their combinations with 4A7 ( D )

Article Snippet: Antibodies used for flow cytometry analysis were mouse anti-HER3 mAb (10,201-MM01) from Sino Biological, Inc. (Beijing, China).

Techniques: Membrane, Activity Assay, Blocking Assay, Migration, Produced, Purification, Binding Assay, Flow Cytometry, Staining, Negative Control, Positive Control, Western Blot, Incubation

Journal: Cancer Cell International

Article Title: HER3 functions as an effective therapeutic target in triple negative breast cancer to potentiate the antitumor activity of gefitinib and paclitaxel

doi: 10.1186/s12935-023-03055-w

Figure Lengend Snippet: Combinations of 4A7 and gefitinib potently inactivate HER3, EGFR, and their downstream signaling, and markedly induce growth inhibition and cell death in TNBC cells. A , The indicated TNBC cell lines cultured at normal condition were collected and subjected to western blot analyses of EGFR, p-EGFR (Y1068), HER3, and p-HER3 (Y1289). β-actin was used as a loading control. The relative signal intensity obtained via densitometry analyses was shown underneath of each image. B , Equal amount of total protein extracts of MDA-MB-468 (MDA-468) or HCC1806 cells were subjected to IP using a control IgG, anti-EGFR Ab (left), or anti-HER3 Ab (right), and followed by western blot analyses of HER3 and EGFR. C , MDA-MB-468 or HCC1806 cells at normal culture condition were treated with vehicle, 4A7, gefitinib, or the combinations of 4A7 and gefitinib for 24 h. The cells were collected and subjected to western blot assays of the indicated markers. Densitometry analyses of the signals were shown underneath, and the arbitrary numbers indicated the intensities of each image relative to the untreated controls, defined as 1.0. D , MDA-MB-468 or HCC1806 cells were plated in 96-well plates. After 24 h, the culture medium was replaced with fresh medium containing vehicle control (DMSO), gefitinib (4µmol/L) alone, or gefitinib (4µmol/L) plus 4A7 (20 µg/ml) (Gefi + 4A7), and incubated for additional 72 h. The percentages of surviving cells relative to controls, defined as 100% survival, were determined by cell growth/viability (MTS) assays. Data shows a representative of three independent experiments. Bars, SD. *, p < 0.05, **, p < 0.01. E , MDA-MB-468 or HCC1806 cells seeded in 6-well plates were treated with gefitinib (8µmol/L) or its combinations with 4A7 (20 µg/ml) for 48 h. The cells were subjected to the LIVE/DEAD Cell Imaging assays. Green, live cells; red, dead cells. The ratio of dead/live cells was shown by in each sample. Data shows a representative of three independent experiments

Article Snippet: Antibodies used for flow cytometry analysis were mouse anti-HER3 mAb (10,201-MM01) from Sino Biological, Inc. (Beijing, China).

Techniques: Inhibition, Cell Culture, Western Blot, Incubation, Imaging

Journal: Cancer Cell International

Article Title: HER3 functions as an effective therapeutic target in triple negative breast cancer to potentiate the antitumor activity of gefitinib and paclitaxel

doi: 10.1186/s12935-023-03055-w

Figure Lengend Snippet: 4A7 significantly enhances paclitaxel-mediated antitumor activity in TNBC tumor xenograft models. HCC1806-luc cells (5 × 10 5 ) were orthotopically inoculated into the mammary fat pads of nude mice to establish tumor xenografts. When tumor volume reached ~ 80mm 3 , the tumor-bearing mice received intraperitoneal injections of PBS, 4A7 (20 mg/kg), paclitaxel (6 mg/kg), or both 4A7 and paclitaxel (Pac + 4A7) (n = 5). After 6 treatments, mice were sacrificed. A , Tumor growth curves were plotted using average tumor volumes within each group at the indicated time points. A two-tailed student’s t-test was used for statistical analysis. Bars, SEM. *, P < 0.01. B , The mammary tumors were dissected and imaged as indicated (Left). The tumors were also measured for weight (Right). Bars, SD. *, p < 0.05, ***, p < 0.005. C and D , At end of the experiments, before mice were sacrificed, bioluminescent imaging of the mammary tumors was obtained with the IVIS spectrum ( C ). The luciferase signal intensity of the mammary tumors from the mice of each treatment group was plotted. Bars, SD. *, p < 0.05, **, p < 0.01. E , Formalin-fixed paraffin-embedded sections of tumors were analyzed with IHC staining for p-HER3 (Y1289), p-EGFR (Y1068), Ki67, or cleaved Caspase-3 (C-caspase-3). Scale bar, 70 μm. Quantification of IHC staining with ImageJ and ImageJ plugin IHC profiler was shown underneath. A two-way ANOVA tests were used for statistical analysis. Bars, SD. *, p < 0.05, ****, p < 0.001

Article Snippet: Antibodies used for flow cytometry analysis were mouse anti-HER3 mAb (10,201-MM01) from Sino Biological, Inc. (Beijing, China).

Techniques: Activity Assay, Two Tailed Test, Imaging, Luciferase, Formalin-fixed Paraffin-Embedded, Immunohistochemistry

Journal: Cancer Cell International

Article Title: HER3 functions as an effective therapeutic target in triple negative breast cancer to potentiate the antitumor activity of gefitinib and paclitaxel

doi: 10.1186/s12935-023-03055-w

Figure Lengend Snippet: Our newly developed anti-HER3 mAb 4A7 recognizes cell membrane HER3, and exhibits profound activity to block HRGβ1-, but not SDF-1-induced TNBC cell migration. A , Our anti-HER3 mAb 4A7 was produced and purified as described in the materials and methods. 4A7 binding to membrane HER3 on SKBR3.B3.1 cells was examined by flow cytometry analysis. Staining with 2nd Ab only was used as a negative control. 10,201-MM01, a commercially available anti-HER3 mAb was used a positive control. Quantification of the positive staining with either 10,201-MM01 or 4A7 from three independent experiments was shown in the bar graphs. B , Equal amount of total protein extracts of HCC1806 cells were either examined by western blot analyses of HER3 and IGF-1R (Input) or subjected to IP (IP: Ab) using a control IgG or 4A7 and followed by western blot analyses of HER3 and IGF-1R. C-D , The specificity of 4A7 against HER3 was confirmed by its capability to block HRGβ1-induced promotion of TNBC cell migration. HCC1806 cells seeded in 96-well plates were pre-incubated with mitomycin C (2 µg/ml) prior to the scratch assays. The cells were then untreated or treated with HRGβ1 (50ng/ml) or HRGβ1 plus 4A7 (20ug/ml) and monitored by an IncuCyte system for cell migration ( C ). The same migration assays were performed with MDA-MB-231 cells in the presence of HRGβ1 (50ng/ml) or SDF-1 (200ng/ml) or their combinations with 4A7 ( D )

Article Snippet: Antibodies used for flow cytometry analysis were mouse anti-HER3 mAb (10,201-MM01) from Sino Biological, Inc. (Beijing, China).

Techniques: Membrane, Activity Assay, Blocking Assay, Migration, Produced, Purification, Binding Assay, Flow Cytometry, Staining, Negative Control, Positive Control, Western Blot, Incubation