rhher3  (R&D Systems)

Journal: International Journal of Molecular Sciences

Article Title: HER4 Affects Sensitivity to Tamoxifen and Abemaciclib in Luminal Breast Cancer Cells and Restricts Tumor Growth in MCF-7-Based Humanized Tumor Mice

doi: 10.3390/ijms25137475

Figure Lengend Snippet: Effects of HER4 knock-out on MCF-7 cells in vitro. ( A ) Long-term treatment of MCF-7 WT (soild blue line) and MCF-7 KO (soild red line) cells with tamoxifen and corresponding cell doubling times (dashed lines) as a function of concentration. ( B ) Abemaciclib long-term treatment of MCF-7 WT and KO cells and corresponding cell doubling times (dashed lines) as a function of concentration. ( C ) Analysis of HER4, 4ICD, and ESR cytoplasmic and nuclear localization by Western blot using fractionated protein lysates (C = cytosol; N = nucleus) of MCF-7 WT cells untreated (ctrl.) and after treatment with tamoxifen (1 µM), E2 (100 nM), or NRG1 (30 ng/mL) for 24 h, respectively. Lamin (nucleus) and tubulin (cytosol) were used as loading controls and indicate the purity of the respective fractions. ( D ) Immunoprecipitation analysis by Western blotting of untreated (ctrl.) and 24 h tamoxifen (1 µM), E2 (100 nM), NRG1 (30 ng/mL), and abemaciclib (100 nM) treated MCF-7 WT cells. Immunoprecipitation was performed using a rabbit anti-HER4 catcher antibody and interaction of HER4/4ICD was analyzed by using a rabbit anti-ESR detection antibody. ( E ) Immunoprecipitation by Western blotting of untreated (ctrl.) and NRG1 (30 ng/mL)-treated MCF-7 WT and KO cells for 6 h. HER3 was detected by using a mouse anti-HER3 antibody upon HER2 immunoprecipitation. ( F ) A supposed model of HER4/4ICD interplay in the presence of E2, tamoxifen, or abemaciclib. Black arrows indicate molecule cross-linking. Red arrows indicate molecule downregulation.

Article Snippet: First, propagated cell clones were analyzed by flow cytometry for HER4 surface expression by a standard staining procedure using a APC-conjugated anti-HER4 (R and D Systems Cat# FAB11311A, RRID:AB_2277999) and corresponding isotype control (BioLegend Cat# 400221, RRID:AB_2891178) antibody.

Techniques: Knock-Out, In Vitro, Concentration Assay, Western Blot, Immunoprecipitation

Journal: International Journal of Molecular Sciences

Article Title: HER4 Affects Sensitivity to Tamoxifen and Abemaciclib in Luminal Breast Cancer Cells and Restricts Tumor Growth in MCF-7-Based Humanized Tumor Mice

doi: 10.3390/ijms25137475

Figure Lengend Snippet: Tumor growth and tumor cell characterization in MCF-7-based HER4 WT and KO HTM. ( A ) Tumor growth of untreated HTM transplanted with MCF-7 WT or KO cells is shown. Data are presented as mean ± SD and two-way ANOVA and Sidak’s multiple comparisons tests were applied. ( B ) Tumor weight at the end of therapy is shown. Two-way ANOVA, Sidak’s, and Dunnett’s multiple comparisons tests were performed. ( C ) Tumor growth of HTM transplanted with MCF-7 WT or KO cells, either untreated (ctrl.) or treated with tamoxifen or abemaciclib, is displayed. Data were analyzed by two-way ANOVA and Dunnett’s multiple comparisons test. ( D ) Single tumor cells were analyzed by flow cytometry and the percentage of HER2 positive cells (of EpCAM pos. cells) is shown. Statistics were performed by unpaired t test, p = 0.0024. Immunohistochemical staining for HER2 was performed and is exemplarily shown for a MCF-7 WT and KO tumor, respectively. ( E ) PD-L1, MHC-I, and MHC-II expression on EPCAM + cells is presented and data were analyzed by one-way ANOVA and Tukey’s multiple comparisons test. ( F ) Tumor cell dissemination in the lung and bone marrow (BM) was assessed by flow cytometry staining of EpCAM positive cells. ( G ) Spleen weight at the end of therapy is shown and differences were analyzed by two-way ANOVA, Sidak’s, and Dunnett’s multiple comparisons tests. ( B , D – G ) Data are shown as mean ± SD and number of animals are indicated by individual symbols. Statistical significances: * p ≤ 0.05, ** p ≤ 0.01 and *** p ≤ 0.001, **** p ≤ 0.0001.

Article Snippet: First, propagated cell clones were analyzed by flow cytometry for HER4 surface expression by a standard staining procedure using a APC-conjugated anti-HER4 (R and D Systems Cat# FAB11311A, RRID:AB_2277999) and corresponding isotype control (BioLegend Cat# 400221, RRID:AB_2891178) antibody.

Techniques: Flow Cytometry, Immunohistochemical staining, Staining, Expressing

Journal: International Journal of Molecular Sciences

Article Title: HER4 Affects Sensitivity to Tamoxifen and Abemaciclib in Luminal Breast Cancer Cells and Restricts Tumor Growth in MCF-7-Based Humanized Tumor Mice

doi: 10.3390/ijms25137475

Figure Lengend Snippet: Event-free survival as a function mdm2 gene copy numbers and HER4 expression. Representative FISH images of mdm2 (green dots) and CEN12 (red dots) without ( A ) and with ( B ) signal gains are shown. ( C ) Kaplan–Meier graph illustrating EFS of luminal BC patients treated with abemaciclib as a function of genomic mdm2 and CEN12. Statistical differences were analyzed by Log-rank (Mantel–Cox) test. ( D – F ) Immunohistochemical HER4 stainings scoring “0” (no expression, “1” (low expression), and “2” (pronounced expression) are exemplified, respectively. ( G ) Kaplan–Meier graphs illustrating EFS of luminal BC patients treated with abemaciclib as a function of HER4 expression. Log-rank (Mantel–Cox) test.

Article Snippet: First, propagated cell clones were analyzed by flow cytometry for HER4 surface expression by a standard staining procedure using a APC-conjugated anti-HER4 (R and D Systems Cat# FAB11311A, RRID:AB_2277999) and corresponding isotype control (BioLegend Cat# 400221, RRID:AB_2891178) antibody.

Techniques: Expressing, Immunohistochemical staining

Journal: International Journal of Molecular Sciences

Article Title: HER4 Affects Sensitivity to Tamoxifen and Abemaciclib in Luminal Breast Cancer Cells and Restricts Tumor Growth in MCF-7-Based Humanized Tumor Mice

doi: 10.3390/ijms25137475

Figure Lengend Snippet: Patients’ demographic data. A total of 39 hormone receptor-positive patients were included into the analyses of mdm2/CEN12 FISH analysis and HER4 IHC. NST = invasive carcinoma of no special type; ET = endocrine therapy; NCT = neoadjuvant chemotherapy; Tx = chemotherapy and/or endocrine therapy.

Article Snippet: First, propagated cell clones were analyzed by flow cytometry for HER4 surface expression by a standard staining procedure using a APC-conjugated anti-HER4 (R and D Systems Cat# FAB11311A, RRID:AB_2277999) and corresponding isotype control (BioLegend Cat# 400221, RRID:AB_2891178) antibody.

Techniques:

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: 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: IC 50 Values of Various Agents on HBCCLs as Assessed by SRB Colorimetric Assay: (A) HER-Family Targeting TKIs and Other Downstream Signaling Molecules and (B) Other TKIs and Chemotherapeutic Agents

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: Colorimetric Assay, Polymer