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p her2 y1248  (R&D Systems)


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    R&D Systems p her2 y1248
    Validation of <t>HER2</t> status and targeted therapy. (A) Immunohistochemistry validation of HER2 staining in the affected lymph node. Scale bars: 2.5 mm, 250 µm, and 100 µm (left to right). (B) Radiologic response of patient. Left, prior to treatment, and right, after four cycles of treatment with T-DM1.
    P Her2 Y1248, supplied by R&D Systems, used in various techniques. Bioz Stars score: 94/100, based on 12 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/product/p+her2+y1248/Human+Phospho-ErbB2%2FHer2+(Y1248)+Antibody/pmc08481915-65-46-48
    Average 94 stars, based on 12 article reviews
    p her2 y1248 - by Bioz Stars, 2026-09
    94/100 stars

    Images

    1) Product Images from "Ex Vivo Drug Screening Informed Targeted Therapy for Metastatic Parotid Squamous Cell Carcinoma"

    Article Title: Ex Vivo Drug Screening Informed Targeted Therapy for Metastatic Parotid Squamous Cell Carcinoma

    Journal: Frontiers in Oncology

    doi: 10.3389/fonc.2021.735820

    Validation of HER2 status and targeted therapy. (A) Immunohistochemistry validation of HER2 staining in the affected lymph node. Scale bars: 2.5 mm, 250 µm, and 100 µm (left to right). (B) Radiologic response of patient. Left, prior to treatment, and right, after four cycles of treatment with T-DM1.
    Figure Legend Snippet: Validation of HER2 status and targeted therapy. (A) Immunohistochemistry validation of HER2 staining in the affected lymph node. Scale bars: 2.5 mm, 250 µm, and 100 µm (left to right). (B) Radiologic response of patient. Left, prior to treatment, and right, after four cycles of treatment with T-DM1.

    Techniques Used: Biomarker Discovery, Immunohistochemistry, Staining

    Repeat ex vivo drug screen in recurrent disease. (A) Scatter plot showing correlation of the mean GR score of 193 drugs in five doses for the HER2 + and VIM + cells. (B) Scatter plot showing correlation of the IC 50 estimate of the analyzed drugs between the HER2 + sample B cells and KRT19 + sample A cells. (C) Box plots showing the GR scores for the five doses of afatinib, neratinib, erlotinib, and lapatinib in sample A and B KRT19 + and HER + cells. Afatinib and neratinib displayed a statistically significant differential effect on the KRT19 + and HER + cells from sample B as compared using paired Student’s t -test. (D) Gallery view of representative 20× immunofluorescence microscopy images of the sample cells treated for 96 h with the different EGFR-TKis in five different concentrations (nM). KRT19 staining shown in blue, VIM staining in green and HER2 staining in red. Bars: 50 µm.
    Figure Legend Snippet: Repeat ex vivo drug screen in recurrent disease. (A) Scatter plot showing correlation of the mean GR score of 193 drugs in five doses for the HER2 + and VIM + cells. (B) Scatter plot showing correlation of the IC 50 estimate of the analyzed drugs between the HER2 + sample B cells and KRT19 + sample A cells. (C) Box plots showing the GR scores for the five doses of afatinib, neratinib, erlotinib, and lapatinib in sample A and B KRT19 + and HER + cells. Afatinib and neratinib displayed a statistically significant differential effect on the KRT19 + and HER + cells from sample B as compared using paired Student’s t -test. (D) Gallery view of representative 20× immunofluorescence microscopy images of the sample cells treated for 96 h with the different EGFR-TKis in five different concentrations (nM). KRT19 staining shown in blue, VIM staining in green and HER2 staining in red. Bars: 50 µm.

    Techniques Used: Ex Vivo, Immunofluorescence, Microscopy, Staining

    A pathway perspective on targeted therapy responses. (A) Heat map visualization with one-dimensional (vertical) unsupervised clustering of the z -scores of the eight RPPA markers across 19 drugs in five concentrations (left to right, low to high, respectively). (B) Top: Representative microscope images of HER2 staining on sample B cells in the ex vivo screen following 96-h exposure to the indicated drugs at 5 µM concentration. KRT19 staining shown in blue, VIM staining in green, and HER2 staining in red. Bars: 50 µm. Bottom left: Box plot showing the mean and standard deviation of fluorescence intensity of HER2 staining (RFU) of the KRT19 + cells in response to treatment with the different EGFR-TKis. Two biological replicate experiments were combined and compared using unpaired Welch’s t -test. Bottom right: Comparison of the HER2 staining intensity to DMSO-treated cells. Signal shown as fold change. (C) Combination of lapatinib and afatanib shows a synergistic growth inhibitory effect. With all three different drug schedules tested, the combination resulted in significantly increased efficacy over the single agents as indicated by the lower IC 50 (middle panel). On the basis of the CI 50 combination index, lapatinib pre-treatment preceding afatinib treatment had the highest synergy (right panel). Comparison was made using unpaired Student’s t -test. (D) The patient-derived tumor cells were treated with DMSO (−), 1000 nmol/L afatinib, lapatinib, neratinib, or combination of 1000 nmol/L afatinib+lapatinib, or neratinib+lapatinib for 48 h, and phosphorylation (p) AKT and total HER2 protein levels of indicated markers were assessed using Western blot. Actin was assessed as a loading control.
    Figure Legend Snippet: A pathway perspective on targeted therapy responses. (A) Heat map visualization with one-dimensional (vertical) unsupervised clustering of the z -scores of the eight RPPA markers across 19 drugs in five concentrations (left to right, low to high, respectively). (B) Top: Representative microscope images of HER2 staining on sample B cells in the ex vivo screen following 96-h exposure to the indicated drugs at 5 µM concentration. KRT19 staining shown in blue, VIM staining in green, and HER2 staining in red. Bars: 50 µm. Bottom left: Box plot showing the mean and standard deviation of fluorescence intensity of HER2 staining (RFU) of the KRT19 + cells in response to treatment with the different EGFR-TKis. Two biological replicate experiments were combined and compared using unpaired Welch’s t -test. Bottom right: Comparison of the HER2 staining intensity to DMSO-treated cells. Signal shown as fold change. (C) Combination of lapatinib and afatanib shows a synergistic growth inhibitory effect. With all three different drug schedules tested, the combination resulted in significantly increased efficacy over the single agents as indicated by the lower IC 50 (middle panel). On the basis of the CI 50 combination index, lapatinib pre-treatment preceding afatinib treatment had the highest synergy (right panel). Comparison was made using unpaired Student’s t -test. (D) The patient-derived tumor cells were treated with DMSO (−), 1000 nmol/L afatinib, lapatinib, neratinib, or combination of 1000 nmol/L afatinib+lapatinib, or neratinib+lapatinib for 48 h, and phosphorylation (p) AKT and total HER2 protein levels of indicated markers were assessed using Western blot. Actin was assessed as a loading control.

    Techniques Used: Microscopy, Staining, Ex Vivo, Concentration Assay, Standard Deviation, Fluorescence, Comparison, Derivative Assay, Phospho-proteomics, Western Blot, Control

    Flow cytometry immunophenotyping of the MISB10B cells. (A) Analysis of expression of epithelial lineage markers and HER2 in the EpCAM + cells. (B) Representative bright-field microscopy image (10×) of the morphology of the MISB10B cells in near full confluent state. (C) Analysis of expression of additional immunophenotypic markers in the EpCAM + cells.
    Figure Legend Snippet: Flow cytometry immunophenotyping of the MISB10B cells. (A) Analysis of expression of epithelial lineage markers and HER2 in the EpCAM + cells. (B) Representative bright-field microscopy image (10×) of the morphology of the MISB10B cells in near full confluent state. (C) Analysis of expression of additional immunophenotypic markers in the EpCAM + cells.

    Techniques Used: Flow Cytometry, Expressing, Microscopy

    Related Articles

    Biomarker Discovery:

    Article Title: Ex Vivo Drug Screening Informed Targeted Therapy for Metastatic Parotid Squamous Cell Carcinoma
    Article Snippet: To generate reverse-phase arrays, lysates were printed on nitrocellulose-coated glass slides (Grace Biolabs #305177) on a Genetix QArray Mini Arrayer (Molecular Devices).To generate reverse-phase arrays, lysates were printed on nitrocellulose-coated glass slides (Grace Biolabs #305177) on a Genetix QArray Mini Arrayer (Molecular Devices).. Primary antibodies used for RPPA experiments contained the following: p-AKT (Ser473) (Cell Signaling Technology Cat# 4060), AKT (Cell Signaling Technology Cat# 4691), p-ERK1/2 (Thr202/Tyr204) (Cell Signaling Technology Cat# 4370), p-S6 Ribosomal protein (Ser235/236) (Cell Signaling Technology Cat# 2211), Ki-67 (Abcam Cat# ab15580), HER2 (DAKO Cat# A0485), p-HER2 (Y1248) (R&D Systems Cat# AF1768), and p-cMET (Tyr1234/1235) (Cell Signaling Technology Cat# 3129).. Secondary detection was performed with a goat anti-mouse IgG conjugated to DyLight 680 (Thermo Pierce Cat# 35518) and goat anti-rabbit IgG conjugated to DyLight 800 (Thermo Pierce Cat# 35571) antibodies.Secondary detection was performed with a goat anti-mouse IgG conjugated to DyLight 680 (Thermo Pierce Cat# 35518) and goat anti-rabbit IgG conjugated to DyLight 800 (Thermo Pierce Cat# 35571) antibodies.

    Article Title: Ex Vivo Drug Screening Informed Targeted Therapy for Metastatic Parotid Squamous Cell Carcinoma.
    Article Snippet: To generate reverse-phase arrays, lysates were printed on nitrocellulose-coated glass slides (Grace Biolabs #305177) on a Genetix QArray Mini Arrayer (Molecular Devices).To generate reverse-phase arrays, lysates were printed on nitrocellulose-coated glass slides (Grace Biolabs #305177) on a Genetix QArray Mini Arrayer (Molecular Devices).. Primary antibodies used for RPPA experiments contained the following: p-AKT (Ser473) (Cell Signaling Technology Cat# 4060), AKT (Cell Signaling Technology Cat# 4691), p-ERK1/2 (Thr202/Tyr204) (Cell Signaling Technology Cat# 4370), p-S6 Ribosomal protein (Ser235/236) (Cell Signaling Technology Cat# 2211), Ki-67 (Abcam Cat# ab15580), HER2 (DAKO Cat# A0485), p-HER2 (Y1248) (R&D Systems Cat# AF1768), and p-cMET (Tyr1234/1235) (Cell Signaling Technology Cat# 3129).. Secondary detection was performed with a goat anti-mouse IgG conjugated to DyLight 680 (Thermo Pierce Cat# 35518) and goat anti-rabbit IgG Frontiers in Oncology | www.frontiersin.org 3 conjugated to DyLight 800 (Thermo Pierce Cat# 35571) antibodies.Secondary detection was performed with a goat anti-mouse IgG conjugated to DyLight 680 (Thermo Pierce Cat# 35518) and goat anti-rabbit IgG Frontiers in Oncology | www.frontiersin.org 3 conjugated to DyLight 800 (Thermo Pierce Cat#..

    Immunohistochemistry:

    Article Title: Ex Vivo Drug Screening Informed Targeted Therapy for Metastatic Parotid Squamous Cell Carcinoma
    Article Snippet: To generate reverse-phase arrays, lysates were printed on nitrocellulose-coated glass slides (Grace Biolabs #305177) on a Genetix QArray Mini Arrayer (Molecular Devices).To generate reverse-phase arrays, lysates were printed on nitrocellulose-coated glass slides (Grace Biolabs #305177) on a Genetix QArray Mini Arrayer (Molecular Devices).. Primary antibodies used for RPPA experiments contained the following: p-AKT (Ser473) (Cell Signaling Technology Cat# 4060), AKT (Cell Signaling Technology Cat# 4691), p-ERK1/2 (Thr202/Tyr204) (Cell Signaling Technology Cat# 4370), p-S6 Ribosomal protein (Ser235/236) (Cell Signaling Technology Cat# 2211), Ki-67 (Abcam Cat# ab15580), HER2 (DAKO Cat# A0485), p-HER2 (Y1248) (R&D Systems Cat# AF1768), and p-cMET (Tyr1234/1235) (Cell Signaling Technology Cat# 3129).. Secondary detection was performed with a goat anti-mouse IgG conjugated to DyLight 680 (Thermo Pierce Cat# 35518) and goat anti-rabbit IgG conjugated to DyLight 800 (Thermo Pierce Cat# 35571) antibodies.Secondary detection was performed with a goat anti-mouse IgG conjugated to DyLight 680 (Thermo Pierce Cat# 35518) and goat anti-rabbit IgG conjugated to DyLight 800 (Thermo Pierce Cat# 35571) antibodies.

    Article Title: Ex Vivo Drug Screening Informed Targeted Therapy for Metastatic Parotid Squamous Cell Carcinoma.
    Article Snippet: To generate reverse-phase arrays, lysates were printed on nitrocellulose-coated glass slides (Grace Biolabs #305177) on a Genetix QArray Mini Arrayer (Molecular Devices).To generate reverse-phase arrays, lysates were printed on nitrocellulose-coated glass slides (Grace Biolabs #305177) on a Genetix QArray Mini Arrayer (Molecular Devices).. Primary antibodies used for RPPA experiments contained the following: p-AKT (Ser473) (Cell Signaling Technology Cat# 4060), AKT (Cell Signaling Technology Cat# 4691), p-ERK1/2 (Thr202/Tyr204) (Cell Signaling Technology Cat# 4370), p-S6 Ribosomal protein (Ser235/236) (Cell Signaling Technology Cat# 2211), Ki-67 (Abcam Cat# ab15580), HER2 (DAKO Cat# A0485), p-HER2 (Y1248) (R&D Systems Cat# AF1768), and p-cMET (Tyr1234/1235) (Cell Signaling Technology Cat# 3129).. Secondary detection was performed with a goat anti-mouse IgG conjugated to DyLight 680 (Thermo Pierce Cat# 35518) and goat anti-rabbit IgG Frontiers in Oncology | www.frontiersin.org 3 conjugated to DyLight 800 (Thermo Pierce Cat# 35571) antibodies.Secondary detection was performed with a goat anti-mouse IgG conjugated to DyLight 680 (Thermo Pierce Cat# 35518) and goat anti-rabbit IgG Frontiers in Oncology | www.frontiersin.org 3 conjugated to DyLight 800 (Thermo Pierce Cat#..

    Staining:

    Article Title: Ex Vivo Drug Screening Informed Targeted Therapy for Metastatic Parotid Squamous Cell Carcinoma
    Article Snippet: To generate reverse-phase arrays, lysates were printed on nitrocellulose-coated glass slides (Grace Biolabs #305177) on a Genetix QArray Mini Arrayer (Molecular Devices).To generate reverse-phase arrays, lysates were printed on nitrocellulose-coated glass slides (Grace Biolabs #305177) on a Genetix QArray Mini Arrayer (Molecular Devices).. Primary antibodies used for RPPA experiments contained the following: p-AKT (Ser473) (Cell Signaling Technology Cat# 4060), AKT (Cell Signaling Technology Cat# 4691), p-ERK1/2 (Thr202/Tyr204) (Cell Signaling Technology Cat# 4370), p-S6 Ribosomal protein (Ser235/236) (Cell Signaling Technology Cat# 2211), Ki-67 (Abcam Cat# ab15580), HER2 (DAKO Cat# A0485), p-HER2 (Y1248) (R&D Systems Cat# AF1768), and p-cMET (Tyr1234/1235) (Cell Signaling Technology Cat# 3129).. Secondary detection was performed with a goat anti-mouse IgG conjugated to DyLight 680 (Thermo Pierce Cat# 35518) and goat anti-rabbit IgG conjugated to DyLight 800 (Thermo Pierce Cat# 35571) antibodies.Secondary detection was performed with a goat anti-mouse IgG conjugated to DyLight 680 (Thermo Pierce Cat# 35518) and goat anti-rabbit IgG conjugated to DyLight 800 (Thermo Pierce Cat# 35571) antibodies.

    Article Title: Ex Vivo Drug Screening Informed Targeted Therapy for Metastatic Parotid Squamous Cell Carcinoma.
    Article Snippet: To generate reverse-phase arrays, lysates were printed on nitrocellulose-coated glass slides (Grace Biolabs #305177) on a Genetix QArray Mini Arrayer (Molecular Devices).To generate reverse-phase arrays, lysates were printed on nitrocellulose-coated glass slides (Grace Biolabs #305177) on a Genetix QArray Mini Arrayer (Molecular Devices).. Primary antibodies used for RPPA experiments contained the following: p-AKT (Ser473) (Cell Signaling Technology Cat# 4060), AKT (Cell Signaling Technology Cat# 4691), p-ERK1/2 (Thr202/Tyr204) (Cell Signaling Technology Cat# 4370), p-S6 Ribosomal protein (Ser235/236) (Cell Signaling Technology Cat# 2211), Ki-67 (Abcam Cat# ab15580), HER2 (DAKO Cat# A0485), p-HER2 (Y1248) (R&D Systems Cat# AF1768), and p-cMET (Tyr1234/1235) (Cell Signaling Technology Cat# 3129).. Secondary detection was performed with a goat anti-mouse IgG conjugated to DyLight 680 (Thermo Pierce Cat# 35518) and goat anti-rabbit IgG Frontiers in Oncology | www.frontiersin.org 3 conjugated to DyLight 800 (Thermo Pierce Cat# 35571) antibodies.Secondary detection was performed with a goat anti-mouse IgG conjugated to DyLight 680 (Thermo Pierce Cat# 35518) and goat anti-rabbit IgG Frontiers in Oncology | www.frontiersin.org 3 conjugated to DyLight 800 (Thermo Pierce Cat#..

    Ex Vivo:

    Article Title: Ex Vivo Drug Screening Informed Targeted Therapy for Metastatic Parotid Squamous Cell Carcinoma
    Article Snippet: To generate reverse-phase arrays, lysates were printed on nitrocellulose-coated glass slides (Grace Biolabs #305177) on a Genetix QArray Mini Arrayer (Molecular Devices).To generate reverse-phase arrays, lysates were printed on nitrocellulose-coated glass slides (Grace Biolabs #305177) on a Genetix QArray Mini Arrayer (Molecular Devices).. Primary antibodies used for RPPA experiments contained the following: p-AKT (Ser473) (Cell Signaling Technology Cat# 4060), AKT (Cell Signaling Technology Cat# 4691), p-ERK1/2 (Thr202/Tyr204) (Cell Signaling Technology Cat# 4370), p-S6 Ribosomal protein (Ser235/236) (Cell Signaling Technology Cat# 2211), Ki-67 (Abcam Cat# ab15580), HER2 (DAKO Cat# A0485), p-HER2 (Y1248) (R&D Systems Cat# AF1768), and p-cMET (Tyr1234/1235) (Cell Signaling Technology Cat# 3129).. Secondary detection was performed with a goat anti-mouse IgG conjugated to DyLight 680 (Thermo Pierce Cat# 35518) and goat anti-rabbit IgG conjugated to DyLight 800 (Thermo Pierce Cat# 35571) antibodies.Secondary detection was performed with a goat anti-mouse IgG conjugated to DyLight 680 (Thermo Pierce Cat# 35518) and goat anti-rabbit IgG conjugated to DyLight 800 (Thermo Pierce Cat# 35571) antibodies.

    Article Title: Ex Vivo Drug Screening Informed Targeted Therapy for Metastatic Parotid Squamous Cell Carcinoma.
    Article Snippet: To generate reverse-phase arrays, lysates were printed on nitrocellulose-coated glass slides (Grace Biolabs #305177) on a Genetix QArray Mini Arrayer (Molecular Devices).To generate reverse-phase arrays, lysates were printed on nitrocellulose-coated glass slides (Grace Biolabs #305177) on a Genetix QArray Mini Arrayer (Molecular Devices).. Primary antibodies used for RPPA experiments contained the following: p-AKT (Ser473) (Cell Signaling Technology Cat# 4060), AKT (Cell Signaling Technology Cat# 4691), p-ERK1/2 (Thr202/Tyr204) (Cell Signaling Technology Cat# 4370), p-S6 Ribosomal protein (Ser235/236) (Cell Signaling Technology Cat# 2211), Ki-67 (Abcam Cat# ab15580), HER2 (DAKO Cat# A0485), p-HER2 (Y1248) (R&D Systems Cat# AF1768), and p-cMET (Tyr1234/1235) (Cell Signaling Technology Cat# 3129).. Secondary detection was performed with a goat anti-mouse IgG conjugated to DyLight 680 (Thermo Pierce Cat# 35518) and goat anti-rabbit IgG Frontiers in Oncology | www.frontiersin.org 3 conjugated to DyLight 800 (Thermo Pierce Cat# 35571) antibodies.Secondary detection was performed with a goat anti-mouse IgG conjugated to DyLight 680 (Thermo Pierce Cat# 35518) and goat anti-rabbit IgG Frontiers in Oncology | www.frontiersin.org 3 conjugated to DyLight 800 (Thermo Pierce Cat#..

    Immunofluorescence:

    Article Title: Ex Vivo Drug Screening Informed Targeted Therapy for Metastatic Parotid Squamous Cell Carcinoma
    Article Snippet: To generate reverse-phase arrays, lysates were printed on nitrocellulose-coated glass slides (Grace Biolabs #305177) on a Genetix QArray Mini Arrayer (Molecular Devices).To generate reverse-phase arrays, lysates were printed on nitrocellulose-coated glass slides (Grace Biolabs #305177) on a Genetix QArray Mini Arrayer (Molecular Devices).. Primary antibodies used for RPPA experiments contained the following: p-AKT (Ser473) (Cell Signaling Technology Cat# 4060), AKT (Cell Signaling Technology Cat# 4691), p-ERK1/2 (Thr202/Tyr204) (Cell Signaling Technology Cat# 4370), p-S6 Ribosomal protein (Ser235/236) (Cell Signaling Technology Cat# 2211), Ki-67 (Abcam Cat# ab15580), HER2 (DAKO Cat# A0485), p-HER2 (Y1248) (R&D Systems Cat# AF1768), and p-cMET (Tyr1234/1235) (Cell Signaling Technology Cat# 3129).. Secondary detection was performed with a goat anti-mouse IgG conjugated to DyLight 680 (Thermo Pierce Cat# 35518) and goat anti-rabbit IgG conjugated to DyLight 800 (Thermo Pierce Cat# 35571) antibodies.Secondary detection was performed with a goat anti-mouse IgG conjugated to DyLight 680 (Thermo Pierce Cat# 35518) and goat anti-rabbit IgG conjugated to DyLight 800 (Thermo Pierce Cat# 35571) antibodies.

    Article Title: Ex Vivo Drug Screening Informed Targeted Therapy for Metastatic Parotid Squamous Cell Carcinoma.
    Article Snippet: To generate reverse-phase arrays, lysates were printed on nitrocellulose-coated glass slides (Grace Biolabs #305177) on a Genetix QArray Mini Arrayer (Molecular Devices).To generate reverse-phase arrays, lysates were printed on nitrocellulose-coated glass slides (Grace Biolabs #305177) on a Genetix QArray Mini Arrayer (Molecular Devices).. Primary antibodies used for RPPA experiments contained the following: p-AKT (Ser473) (Cell Signaling Technology Cat# 4060), AKT (Cell Signaling Technology Cat# 4691), p-ERK1/2 (Thr202/Tyr204) (Cell Signaling Technology Cat# 4370), p-S6 Ribosomal protein (Ser235/236) (Cell Signaling Technology Cat# 2211), Ki-67 (Abcam Cat# ab15580), HER2 (DAKO Cat# A0485), p-HER2 (Y1248) (R&D Systems Cat# AF1768), and p-cMET (Tyr1234/1235) (Cell Signaling Technology Cat# 3129).. Secondary detection was performed with a goat anti-mouse IgG conjugated to DyLight 680 (Thermo Pierce Cat# 35518) and goat anti-rabbit IgG Frontiers in Oncology | www.frontiersin.org 3 conjugated to DyLight 800 (Thermo Pierce Cat# 35571) antibodies.Secondary detection was performed with a goat anti-mouse IgG conjugated to DyLight 680 (Thermo Pierce Cat# 35518) and goat anti-rabbit IgG Frontiers in Oncology | www.frontiersin.org 3 conjugated to DyLight 800 (Thermo Pierce Cat#..

    Microscopy:

    Article Title: Ex Vivo Drug Screening Informed Targeted Therapy for Metastatic Parotid Squamous Cell Carcinoma
    Article Snippet: To generate reverse-phase arrays, lysates were printed on nitrocellulose-coated glass slides (Grace Biolabs #305177) on a Genetix QArray Mini Arrayer (Molecular Devices).To generate reverse-phase arrays, lysates were printed on nitrocellulose-coated glass slides (Grace Biolabs #305177) on a Genetix QArray Mini Arrayer (Molecular Devices).. Primary antibodies used for RPPA experiments contained the following: p-AKT (Ser473) (Cell Signaling Technology Cat# 4060), AKT (Cell Signaling Technology Cat# 4691), p-ERK1/2 (Thr202/Tyr204) (Cell Signaling Technology Cat# 4370), p-S6 Ribosomal protein (Ser235/236) (Cell Signaling Technology Cat# 2211), Ki-67 (Abcam Cat# ab15580), HER2 (DAKO Cat# A0485), p-HER2 (Y1248) (R&D Systems Cat# AF1768), and p-cMET (Tyr1234/1235) (Cell Signaling Technology Cat# 3129).. Secondary detection was performed with a goat anti-mouse IgG conjugated to DyLight 680 (Thermo Pierce Cat# 35518) and goat anti-rabbit IgG conjugated to DyLight 800 (Thermo Pierce Cat# 35571) antibodies.Secondary detection was performed with a goat anti-mouse IgG conjugated to DyLight 680 (Thermo Pierce Cat# 35518) and goat anti-rabbit IgG conjugated to DyLight 800 (Thermo Pierce Cat# 35571) antibodies.

    Article Title: Ex Vivo Drug Screening Informed Targeted Therapy for Metastatic Parotid Squamous Cell Carcinoma.
    Article Snippet: To generate reverse-phase arrays, lysates were printed on nitrocellulose-coated glass slides (Grace Biolabs #305177) on a Genetix QArray Mini Arrayer (Molecular Devices).To generate reverse-phase arrays, lysates were printed on nitrocellulose-coated glass slides (Grace Biolabs #305177) on a Genetix QArray Mini Arrayer (Molecular Devices).. Primary antibodies used for RPPA experiments contained the following: p-AKT (Ser473) (Cell Signaling Technology Cat# 4060), AKT (Cell Signaling Technology Cat# 4691), p-ERK1/2 (Thr202/Tyr204) (Cell Signaling Technology Cat# 4370), p-S6 Ribosomal protein (Ser235/236) (Cell Signaling Technology Cat# 2211), Ki-67 (Abcam Cat# ab15580), HER2 (DAKO Cat# A0485), p-HER2 (Y1248) (R&D Systems Cat# AF1768), and p-cMET (Tyr1234/1235) (Cell Signaling Technology Cat# 3129).. Secondary detection was performed with a goat anti-mouse IgG conjugated to DyLight 680 (Thermo Pierce Cat# 35518) and goat anti-rabbit IgG Frontiers in Oncology | www.frontiersin.org 3 conjugated to DyLight 800 (Thermo Pierce Cat# 35571) antibodies.Secondary detection was performed with a goat anti-mouse IgG conjugated to DyLight 680 (Thermo Pierce Cat# 35518) and goat anti-rabbit IgG Frontiers in Oncology | www.frontiersin.org 3 conjugated to DyLight 800 (Thermo Pierce Cat#..

    Concentration Assay:

    Article Title: Ex Vivo Drug Screening Informed Targeted Therapy for Metastatic Parotid Squamous Cell Carcinoma
    Article Snippet: To generate reverse-phase arrays, lysates were printed on nitrocellulose-coated glass slides (Grace Biolabs #305177) on a Genetix QArray Mini Arrayer (Molecular Devices).To generate reverse-phase arrays, lysates were printed on nitrocellulose-coated glass slides (Grace Biolabs #305177) on a Genetix QArray Mini Arrayer (Molecular Devices).. Primary antibodies used for RPPA experiments contained the following: p-AKT (Ser473) (Cell Signaling Technology Cat# 4060), AKT (Cell Signaling Technology Cat# 4691), p-ERK1/2 (Thr202/Tyr204) (Cell Signaling Technology Cat# 4370), p-S6 Ribosomal protein (Ser235/236) (Cell Signaling Technology Cat# 2211), Ki-67 (Abcam Cat# ab15580), HER2 (DAKO Cat# A0485), p-HER2 (Y1248) (R&D Systems Cat# AF1768), and p-cMET (Tyr1234/1235) (Cell Signaling Technology Cat# 3129).. Secondary detection was performed with a goat anti-mouse IgG conjugated to DyLight 680 (Thermo Pierce Cat# 35518) and goat anti-rabbit IgG conjugated to DyLight 800 (Thermo Pierce Cat# 35571) antibodies.Secondary detection was performed with a goat anti-mouse IgG conjugated to DyLight 680 (Thermo Pierce Cat# 35518) and goat anti-rabbit IgG conjugated to DyLight 800 (Thermo Pierce Cat# 35571) antibodies.

    Article Title: Ex Vivo Drug Screening Informed Targeted Therapy for Metastatic Parotid Squamous Cell Carcinoma.
    Article Snippet: To generate reverse-phase arrays, lysates were printed on nitrocellulose-coated glass slides (Grace Biolabs #305177) on a Genetix QArray Mini Arrayer (Molecular Devices).To generate reverse-phase arrays, lysates were printed on nitrocellulose-coated glass slides (Grace Biolabs #305177) on a Genetix QArray Mini Arrayer (Molecular Devices).. Primary antibodies used for RPPA experiments contained the following: p-AKT (Ser473) (Cell Signaling Technology Cat# 4060), AKT (Cell Signaling Technology Cat# 4691), p-ERK1/2 (Thr202/Tyr204) (Cell Signaling Technology Cat# 4370), p-S6 Ribosomal protein (Ser235/236) (Cell Signaling Technology Cat# 2211), Ki-67 (Abcam Cat# ab15580), HER2 (DAKO Cat# A0485), p-HER2 (Y1248) (R&D Systems Cat# AF1768), and p-cMET (Tyr1234/1235) (Cell Signaling Technology Cat# 3129).. Secondary detection was performed with a goat anti-mouse IgG conjugated to DyLight 680 (Thermo Pierce Cat# 35518) and goat anti-rabbit IgG Frontiers in Oncology | www.frontiersin.org 3 conjugated to DyLight 800 (Thermo Pierce Cat# 35571) antibodies.Secondary detection was performed with a goat anti-mouse IgG conjugated to DyLight 680 (Thermo Pierce Cat# 35518) and goat anti-rabbit IgG Frontiers in Oncology | www.frontiersin.org 3 conjugated to DyLight 800 (Thermo Pierce Cat#..

    Standard Deviation:

    Article Title: Ex Vivo Drug Screening Informed Targeted Therapy for Metastatic Parotid Squamous Cell Carcinoma
    Article Snippet: To generate reverse-phase arrays, lysates were printed on nitrocellulose-coated glass slides (Grace Biolabs #305177) on a Genetix QArray Mini Arrayer (Molecular Devices).To generate reverse-phase arrays, lysates were printed on nitrocellulose-coated glass slides (Grace Biolabs #305177) on a Genetix QArray Mini Arrayer (Molecular Devices).. Primary antibodies used for RPPA experiments contained the following: p-AKT (Ser473) (Cell Signaling Technology Cat# 4060), AKT (Cell Signaling Technology Cat# 4691), p-ERK1/2 (Thr202/Tyr204) (Cell Signaling Technology Cat# 4370), p-S6 Ribosomal protein (Ser235/236) (Cell Signaling Technology Cat# 2211), Ki-67 (Abcam Cat# ab15580), HER2 (DAKO Cat# A0485), p-HER2 (Y1248) (R&D Systems Cat# AF1768), and p-cMET (Tyr1234/1235) (Cell Signaling Technology Cat# 3129).. Secondary detection was performed with a goat anti-mouse IgG conjugated to DyLight 680 (Thermo Pierce Cat# 35518) and goat anti-rabbit IgG conjugated to DyLight 800 (Thermo Pierce Cat# 35571) antibodies.Secondary detection was performed with a goat anti-mouse IgG conjugated to DyLight 680 (Thermo Pierce Cat# 35518) and goat anti-rabbit IgG conjugated to DyLight 800 (Thermo Pierce Cat# 35571) antibodies.

    Article Title: Ex Vivo Drug Screening Informed Targeted Therapy for Metastatic Parotid Squamous Cell Carcinoma.
    Article Snippet: To generate reverse-phase arrays, lysates were printed on nitrocellulose-coated glass slides (Grace Biolabs #305177) on a Genetix QArray Mini Arrayer (Molecular Devices).To generate reverse-phase arrays, lysates were printed on nitrocellulose-coated glass slides (Grace Biolabs #305177) on a Genetix QArray Mini Arrayer (Molecular Devices).. Primary antibodies used for RPPA experiments contained the following: p-AKT (Ser473) (Cell Signaling Technology Cat# 4060), AKT (Cell Signaling Technology Cat# 4691), p-ERK1/2 (Thr202/Tyr204) (Cell Signaling Technology Cat# 4370), p-S6 Ribosomal protein (Ser235/236) (Cell Signaling Technology Cat# 2211), Ki-67 (Abcam Cat# ab15580), HER2 (DAKO Cat# A0485), p-HER2 (Y1248) (R&D Systems Cat# AF1768), and p-cMET (Tyr1234/1235) (Cell Signaling Technology Cat# 3129).. Secondary detection was performed with a goat anti-mouse IgG conjugated to DyLight 680 (Thermo Pierce Cat# 35518) and goat anti-rabbit IgG Frontiers in Oncology | www.frontiersin.org 3 conjugated to DyLight 800 (Thermo Pierce Cat# 35571) antibodies.Secondary detection was performed with a goat anti-mouse IgG conjugated to DyLight 680 (Thermo Pierce Cat# 35518) and goat anti-rabbit IgG Frontiers in Oncology | www.frontiersin.org 3 conjugated to DyLight 800 (Thermo Pierce Cat#..

    Fluorescence:

    Article Title: Ex Vivo Drug Screening Informed Targeted Therapy for Metastatic Parotid Squamous Cell Carcinoma
    Article Snippet: To generate reverse-phase arrays, lysates were printed on nitrocellulose-coated glass slides (Grace Biolabs #305177) on a Genetix QArray Mini Arrayer (Molecular Devices).To generate reverse-phase arrays, lysates were printed on nitrocellulose-coated glass slides (Grace Biolabs #305177) on a Genetix QArray Mini Arrayer (Molecular Devices).. Primary antibodies used for RPPA experiments contained the following: p-AKT (Ser473) (Cell Signaling Technology Cat# 4060), AKT (Cell Signaling Technology Cat# 4691), p-ERK1/2 (Thr202/Tyr204) (Cell Signaling Technology Cat# 4370), p-S6 Ribosomal protein (Ser235/236) (Cell Signaling Technology Cat# 2211), Ki-67 (Abcam Cat# ab15580), HER2 (DAKO Cat# A0485), p-HER2 (Y1248) (R&D Systems Cat# AF1768), and p-cMET (Tyr1234/1235) (Cell Signaling Technology Cat# 3129).. Secondary detection was performed with a goat anti-mouse IgG conjugated to DyLight 680 (Thermo Pierce Cat# 35518) and goat anti-rabbit IgG conjugated to DyLight 800 (Thermo Pierce Cat# 35571) antibodies.Secondary detection was performed with a goat anti-mouse IgG conjugated to DyLight 680 (Thermo Pierce Cat# 35518) and goat anti-rabbit IgG conjugated to DyLight 800 (Thermo Pierce Cat# 35571) antibodies.

    Article Title: Ex Vivo Drug Screening Informed Targeted Therapy for Metastatic Parotid Squamous Cell Carcinoma.
    Article Snippet: To generate reverse-phase arrays, lysates were printed on nitrocellulose-coated glass slides (Grace Biolabs #305177) on a Genetix QArray Mini Arrayer (Molecular Devices).To generate reverse-phase arrays, lysates were printed on nitrocellulose-coated glass slides (Grace Biolabs #305177) on a Genetix QArray Mini Arrayer (Molecular Devices).. Primary antibodies used for RPPA experiments contained the following: p-AKT (Ser473) (Cell Signaling Technology Cat# 4060), AKT (Cell Signaling Technology Cat# 4691), p-ERK1/2 (Thr202/Tyr204) (Cell Signaling Technology Cat# 4370), p-S6 Ribosomal protein (Ser235/236) (Cell Signaling Technology Cat# 2211), Ki-67 (Abcam Cat# ab15580), HER2 (DAKO Cat# A0485), p-HER2 (Y1248) (R&D Systems Cat# AF1768), and p-cMET (Tyr1234/1235) (Cell Signaling Technology Cat# 3129).. Secondary detection was performed with a goat anti-mouse IgG conjugated to DyLight 680 (Thermo Pierce Cat# 35518) and goat anti-rabbit IgG Frontiers in Oncology | www.frontiersin.org 3 conjugated to DyLight 800 (Thermo Pierce Cat# 35571) antibodies.Secondary detection was performed with a goat anti-mouse IgG conjugated to DyLight 680 (Thermo Pierce Cat# 35518) and goat anti-rabbit IgG Frontiers in Oncology | www.frontiersin.org 3 conjugated to DyLight 800 (Thermo Pierce Cat#..

    Comparison:

    Article Title: Ex Vivo Drug Screening Informed Targeted Therapy for Metastatic Parotid Squamous Cell Carcinoma
    Article Snippet: To generate reverse-phase arrays, lysates were printed on nitrocellulose-coated glass slides (Grace Biolabs #305177) on a Genetix QArray Mini Arrayer (Molecular Devices).To generate reverse-phase arrays, lysates were printed on nitrocellulose-coated glass slides (Grace Biolabs #305177) on a Genetix QArray Mini Arrayer (Molecular Devices).. Primary antibodies used for RPPA experiments contained the following: p-AKT (Ser473) (Cell Signaling Technology Cat# 4060), AKT (Cell Signaling Technology Cat# 4691), p-ERK1/2 (Thr202/Tyr204) (Cell Signaling Technology Cat# 4370), p-S6 Ribosomal protein (Ser235/236) (Cell Signaling Technology Cat# 2211), Ki-67 (Abcam Cat# ab15580), HER2 (DAKO Cat# A0485), p-HER2 (Y1248) (R&D Systems Cat# AF1768), and p-cMET (Tyr1234/1235) (Cell Signaling Technology Cat# 3129).. Secondary detection was performed with a goat anti-mouse IgG conjugated to DyLight 680 (Thermo Pierce Cat# 35518) and goat anti-rabbit IgG conjugated to DyLight 800 (Thermo Pierce Cat# 35571) antibodies.Secondary detection was performed with a goat anti-mouse IgG conjugated to DyLight 680 (Thermo Pierce Cat# 35518) and goat anti-rabbit IgG conjugated to DyLight 800 (Thermo Pierce Cat# 35571) antibodies.

    Article Title: Ex Vivo Drug Screening Informed Targeted Therapy for Metastatic Parotid Squamous Cell Carcinoma.
    Article Snippet: To generate reverse-phase arrays, lysates were printed on nitrocellulose-coated glass slides (Grace Biolabs #305177) on a Genetix QArray Mini Arrayer (Molecular Devices).To generate reverse-phase arrays, lysates were printed on nitrocellulose-coated glass slides (Grace Biolabs #305177) on a Genetix QArray Mini Arrayer (Molecular Devices).. Primary antibodies used for RPPA experiments contained the following: p-AKT (Ser473) (Cell Signaling Technology Cat# 4060), AKT (Cell Signaling Technology Cat# 4691), p-ERK1/2 (Thr202/Tyr204) (Cell Signaling Technology Cat# 4370), p-S6 Ribosomal protein (Ser235/236) (Cell Signaling Technology Cat# 2211), Ki-67 (Abcam Cat# ab15580), HER2 (DAKO Cat# A0485), p-HER2 (Y1248) (R&D Systems Cat# AF1768), and p-cMET (Tyr1234/1235) (Cell Signaling Technology Cat# 3129).. Secondary detection was performed with a goat anti-mouse IgG conjugated to DyLight 680 (Thermo Pierce Cat# 35518) and goat anti-rabbit IgG Frontiers in Oncology | www.frontiersin.org 3 conjugated to DyLight 800 (Thermo Pierce Cat# 35571) antibodies.Secondary detection was performed with a goat anti-mouse IgG conjugated to DyLight 680 (Thermo Pierce Cat# 35518) and goat anti-rabbit IgG Frontiers in Oncology | www.frontiersin.org 3 conjugated to DyLight 800 (Thermo Pierce Cat#..

    Derivative Assay:

    Article Title: Ex Vivo Drug Screening Informed Targeted Therapy for Metastatic Parotid Squamous Cell Carcinoma
    Article Snippet: To generate reverse-phase arrays, lysates were printed on nitrocellulose-coated glass slides (Grace Biolabs #305177) on a Genetix QArray Mini Arrayer (Molecular Devices).To generate reverse-phase arrays, lysates were printed on nitrocellulose-coated glass slides (Grace Biolabs #305177) on a Genetix QArray Mini Arrayer (Molecular Devices).. Primary antibodies used for RPPA experiments contained the following: p-AKT (Ser473) (Cell Signaling Technology Cat# 4060), AKT (Cell Signaling Technology Cat# 4691), p-ERK1/2 (Thr202/Tyr204) (Cell Signaling Technology Cat# 4370), p-S6 Ribosomal protein (Ser235/236) (Cell Signaling Technology Cat# 2211), Ki-67 (Abcam Cat# ab15580), HER2 (DAKO Cat# A0485), p-HER2 (Y1248) (R&D Systems Cat# AF1768), and p-cMET (Tyr1234/1235) (Cell Signaling Technology Cat# 3129).. Secondary detection was performed with a goat anti-mouse IgG conjugated to DyLight 680 (Thermo Pierce Cat# 35518) and goat anti-rabbit IgG conjugated to DyLight 800 (Thermo Pierce Cat# 35571) antibodies.Secondary detection was performed with a goat anti-mouse IgG conjugated to DyLight 680 (Thermo Pierce Cat# 35518) and goat anti-rabbit IgG conjugated to DyLight 800 (Thermo Pierce Cat# 35571) antibodies.

    Article Title: Ex Vivo Drug Screening Informed Targeted Therapy for Metastatic Parotid Squamous Cell Carcinoma.
    Article Snippet: To generate reverse-phase arrays, lysates were printed on nitrocellulose-coated glass slides (Grace Biolabs #305177) on a Genetix QArray Mini Arrayer (Molecular Devices).To generate reverse-phase arrays, lysates were printed on nitrocellulose-coated glass slides (Grace Biolabs #305177) on a Genetix QArray Mini Arrayer (Molecular Devices).. Primary antibodies used for RPPA experiments contained the following: p-AKT (Ser473) (Cell Signaling Technology Cat# 4060), AKT (Cell Signaling Technology Cat# 4691), p-ERK1/2 (Thr202/Tyr204) (Cell Signaling Technology Cat# 4370), p-S6 Ribosomal protein (Ser235/236) (Cell Signaling Technology Cat# 2211), Ki-67 (Abcam Cat# ab15580), HER2 (DAKO Cat# A0485), p-HER2 (Y1248) (R&D Systems Cat# AF1768), and p-cMET (Tyr1234/1235) (Cell Signaling Technology Cat# 3129).. Secondary detection was performed with a goat anti-mouse IgG conjugated to DyLight 680 (Thermo Pierce Cat# 35518) and goat anti-rabbit IgG Frontiers in Oncology | www.frontiersin.org 3 conjugated to DyLight 800 (Thermo Pierce Cat# 35571) antibodies.Secondary detection was performed with a goat anti-mouse IgG conjugated to DyLight 680 (Thermo Pierce Cat# 35518) and goat anti-rabbit IgG Frontiers in Oncology | www.frontiersin.org 3 conjugated to DyLight 800 (Thermo Pierce Cat#..

    Phospho-proteomics:

    Article Title: Ex Vivo Drug Screening Informed Targeted Therapy for Metastatic Parotid Squamous Cell Carcinoma
    Article Snippet: To generate reverse-phase arrays, lysates were printed on nitrocellulose-coated glass slides (Grace Biolabs #305177) on a Genetix QArray Mini Arrayer (Molecular Devices).To generate reverse-phase arrays, lysates were printed on nitrocellulose-coated glass slides (Grace Biolabs #305177) on a Genetix QArray Mini Arrayer (Molecular Devices).. Primary antibodies used for RPPA experiments contained the following: p-AKT (Ser473) (Cell Signaling Technology Cat# 4060), AKT (Cell Signaling Technology Cat# 4691), p-ERK1/2 (Thr202/Tyr204) (Cell Signaling Technology Cat# 4370), p-S6 Ribosomal protein (Ser235/236) (Cell Signaling Technology Cat# 2211), Ki-67 (Abcam Cat# ab15580), HER2 (DAKO Cat# A0485), p-HER2 (Y1248) (R&D Systems Cat# AF1768), and p-cMET (Tyr1234/1235) (Cell Signaling Technology Cat# 3129).. Secondary detection was performed with a goat anti-mouse IgG conjugated to DyLight 680 (Thermo Pierce Cat# 35518) and goat anti-rabbit IgG conjugated to DyLight 800 (Thermo Pierce Cat# 35571) antibodies.Secondary detection was performed with a goat anti-mouse IgG conjugated to DyLight 680 (Thermo Pierce Cat# 35518) and goat anti-rabbit IgG conjugated to DyLight 800 (Thermo Pierce Cat# 35571) antibodies.

    Article Title: Ex Vivo Drug Screening Informed Targeted Therapy for Metastatic Parotid Squamous Cell Carcinoma.
    Article Snippet: To generate reverse-phase arrays, lysates were printed on nitrocellulose-coated glass slides (Grace Biolabs #305177) on a Genetix QArray Mini Arrayer (Molecular Devices).To generate reverse-phase arrays, lysates were printed on nitrocellulose-coated glass slides (Grace Biolabs #305177) on a Genetix QArray Mini Arrayer (Molecular Devices).. Primary antibodies used for RPPA experiments contained the following: p-AKT (Ser473) (Cell Signaling Technology Cat# 4060), AKT (Cell Signaling Technology Cat# 4691), p-ERK1/2 (Thr202/Tyr204) (Cell Signaling Technology Cat# 4370), p-S6 Ribosomal protein (Ser235/236) (Cell Signaling Technology Cat# 2211), Ki-67 (Abcam Cat# ab15580), HER2 (DAKO Cat# A0485), p-HER2 (Y1248) (R&D Systems Cat# AF1768), and p-cMET (Tyr1234/1235) (Cell Signaling Technology Cat# 3129).. Secondary detection was performed with a goat anti-mouse IgG conjugated to DyLight 680 (Thermo Pierce Cat# 35518) and goat anti-rabbit IgG Frontiers in Oncology | www.frontiersin.org 3 conjugated to DyLight 800 (Thermo Pierce Cat# 35571) antibodies.Secondary detection was performed with a goat anti-mouse IgG conjugated to DyLight 680 (Thermo Pierce Cat# 35518) and goat anti-rabbit IgG Frontiers in Oncology | www.frontiersin.org 3 conjugated to DyLight 800 (Thermo Pierce Cat#..

    Western Blot:

    Article Title: Ex Vivo Drug Screening Informed Targeted Therapy for Metastatic Parotid Squamous Cell Carcinoma
    Article Snippet: To generate reverse-phase arrays, lysates were printed on nitrocellulose-coated glass slides (Grace Biolabs #305177) on a Genetix QArray Mini Arrayer (Molecular Devices).To generate reverse-phase arrays, lysates were printed on nitrocellulose-coated glass slides (Grace Biolabs #305177) on a Genetix QArray Mini Arrayer (Molecular Devices).. Primary antibodies used for RPPA experiments contained the following: p-AKT (Ser473) (Cell Signaling Technology Cat# 4060), AKT (Cell Signaling Technology Cat# 4691), p-ERK1/2 (Thr202/Tyr204) (Cell Signaling Technology Cat# 4370), p-S6 Ribosomal protein (Ser235/236) (Cell Signaling Technology Cat# 2211), Ki-67 (Abcam Cat# ab15580), HER2 (DAKO Cat# A0485), p-HER2 (Y1248) (R&D Systems Cat# AF1768), and p-cMET (Tyr1234/1235) (Cell Signaling Technology Cat# 3129).. Secondary detection was performed with a goat anti-mouse IgG conjugated to DyLight 680 (Thermo Pierce Cat# 35518) and goat anti-rabbit IgG conjugated to DyLight 800 (Thermo Pierce Cat# 35571) antibodies.Secondary detection was performed with a goat anti-mouse IgG conjugated to DyLight 680 (Thermo Pierce Cat# 35518) and goat anti-rabbit IgG conjugated to DyLight 800 (Thermo Pierce Cat# 35571) antibodies.

    Article Title: Ex Vivo Drug Screening Informed Targeted Therapy for Metastatic Parotid Squamous Cell Carcinoma.
    Article Snippet: To generate reverse-phase arrays, lysates were printed on nitrocellulose-coated glass slides (Grace Biolabs #305177) on a Genetix QArray Mini Arrayer (Molecular Devices).To generate reverse-phase arrays, lysates were printed on nitrocellulose-coated glass slides (Grace Biolabs #305177) on a Genetix QArray Mini Arrayer (Molecular Devices).. Primary antibodies used for RPPA experiments contained the following: p-AKT (Ser473) (Cell Signaling Technology Cat# 4060), AKT (Cell Signaling Technology Cat# 4691), p-ERK1/2 (Thr202/Tyr204) (Cell Signaling Technology Cat# 4370), p-S6 Ribosomal protein (Ser235/236) (Cell Signaling Technology Cat# 2211), Ki-67 (Abcam Cat# ab15580), HER2 (DAKO Cat# A0485), p-HER2 (Y1248) (R&D Systems Cat# AF1768), and p-cMET (Tyr1234/1235) (Cell Signaling Technology Cat# 3129).. Secondary detection was performed with a goat anti-mouse IgG conjugated to DyLight 680 (Thermo Pierce Cat# 35518) and goat anti-rabbit IgG Frontiers in Oncology | www.frontiersin.org 3 conjugated to DyLight 800 (Thermo Pierce Cat# 35571) antibodies.Secondary detection was performed with a goat anti-mouse IgG conjugated to DyLight 680 (Thermo Pierce Cat# 35518) and goat anti-rabbit IgG Frontiers in Oncology | www.frontiersin.org 3 conjugated to DyLight 800 (Thermo Pierce Cat#..

    Control:

    Article Title: Ex Vivo Drug Screening Informed Targeted Therapy for Metastatic Parotid Squamous Cell Carcinoma
    Article Snippet: To generate reverse-phase arrays, lysates were printed on nitrocellulose-coated glass slides (Grace Biolabs #305177) on a Genetix QArray Mini Arrayer (Molecular Devices).To generate reverse-phase arrays, lysates were printed on nitrocellulose-coated glass slides (Grace Biolabs #305177) on a Genetix QArray Mini Arrayer (Molecular Devices).. Primary antibodies used for RPPA experiments contained the following: p-AKT (Ser473) (Cell Signaling Technology Cat# 4060), AKT (Cell Signaling Technology Cat# 4691), p-ERK1/2 (Thr202/Tyr204) (Cell Signaling Technology Cat# 4370), p-S6 Ribosomal protein (Ser235/236) (Cell Signaling Technology Cat# 2211), Ki-67 (Abcam Cat# ab15580), HER2 (DAKO Cat# A0485), p-HER2 (Y1248) (R&D Systems Cat# AF1768), and p-cMET (Tyr1234/1235) (Cell Signaling Technology Cat# 3129).. Secondary detection was performed with a goat anti-mouse IgG conjugated to DyLight 680 (Thermo Pierce Cat# 35518) and goat anti-rabbit IgG conjugated to DyLight 800 (Thermo Pierce Cat# 35571) antibodies.Secondary detection was performed with a goat anti-mouse IgG conjugated to DyLight 680 (Thermo Pierce Cat# 35518) and goat anti-rabbit IgG conjugated to DyLight 800 (Thermo Pierce Cat# 35571) antibodies.

    Article Title: Ex Vivo Drug Screening Informed Targeted Therapy for Metastatic Parotid Squamous Cell Carcinoma.
    Article Snippet: To generate reverse-phase arrays, lysates were printed on nitrocellulose-coated glass slides (Grace Biolabs #305177) on a Genetix QArray Mini Arrayer (Molecular Devices).To generate reverse-phase arrays, lysates were printed on nitrocellulose-coated glass slides (Grace Biolabs #305177) on a Genetix QArray Mini Arrayer (Molecular Devices).. Primary antibodies used for RPPA experiments contained the following: p-AKT (Ser473) (Cell Signaling Technology Cat# 4060), AKT (Cell Signaling Technology Cat# 4691), p-ERK1/2 (Thr202/Tyr204) (Cell Signaling Technology Cat# 4370), p-S6 Ribosomal protein (Ser235/236) (Cell Signaling Technology Cat# 2211), Ki-67 (Abcam Cat# ab15580), HER2 (DAKO Cat# A0485), p-HER2 (Y1248) (R&D Systems Cat# AF1768), and p-cMET (Tyr1234/1235) (Cell Signaling Technology Cat# 3129).. Secondary detection was performed with a goat anti-mouse IgG conjugated to DyLight 680 (Thermo Pierce Cat# 35518) and goat anti-rabbit IgG Frontiers in Oncology | www.frontiersin.org 3 conjugated to DyLight 800 (Thermo Pierce Cat# 35571) antibodies.Secondary detection was performed with a goat anti-mouse IgG conjugated to DyLight 680 (Thermo Pierce Cat# 35518) and goat anti-rabbit IgG Frontiers in Oncology | www.frontiersin.org 3 conjugated to DyLight 800 (Thermo Pierce Cat#..

    Flow Cytometry:

    Article Title: Ex Vivo Drug Screening Informed Targeted Therapy for Metastatic Parotid Squamous Cell Carcinoma
    Article Snippet: To generate reverse-phase arrays, lysates were printed on nitrocellulose-coated glass slides (Grace Biolabs #305177) on a Genetix QArray Mini Arrayer (Molecular Devices).To generate reverse-phase arrays, lysates were printed on nitrocellulose-coated glass slides (Grace Biolabs #305177) on a Genetix QArray Mini Arrayer (Molecular Devices).. Primary antibodies used for RPPA experiments contained the following: p-AKT (Ser473) (Cell Signaling Technology Cat# 4060), AKT (Cell Signaling Technology Cat# 4691), p-ERK1/2 (Thr202/Tyr204) (Cell Signaling Technology Cat# 4370), p-S6 Ribosomal protein (Ser235/236) (Cell Signaling Technology Cat# 2211), Ki-67 (Abcam Cat# ab15580), HER2 (DAKO Cat# A0485), p-HER2 (Y1248) (R&D Systems Cat# AF1768), and p-cMET (Tyr1234/1235) (Cell Signaling Technology Cat# 3129).. Secondary detection was performed with a goat anti-mouse IgG conjugated to DyLight 680 (Thermo Pierce Cat# 35518) and goat anti-rabbit IgG conjugated to DyLight 800 (Thermo Pierce Cat# 35571) antibodies.Secondary detection was performed with a goat anti-mouse IgG conjugated to DyLight 680 (Thermo Pierce Cat# 35518) and goat anti-rabbit IgG conjugated to DyLight 800 (Thermo Pierce Cat# 35571) antibodies.

    Article Title: Ex Vivo Drug Screening Informed Targeted Therapy for Metastatic Parotid Squamous Cell Carcinoma.
    Article Snippet: To generate reverse-phase arrays, lysates were printed on nitrocellulose-coated glass slides (Grace Biolabs #305177) on a Genetix QArray Mini Arrayer (Molecular Devices).To generate reverse-phase arrays, lysates were printed on nitrocellulose-coated glass slides (Grace Biolabs #305177) on a Genetix QArray Mini Arrayer (Molecular Devices).. Primary antibodies used for RPPA experiments contained the following: p-AKT (Ser473) (Cell Signaling Technology Cat# 4060), AKT (Cell Signaling Technology Cat# 4691), p-ERK1/2 (Thr202/Tyr204) (Cell Signaling Technology Cat# 4370), p-S6 Ribosomal protein (Ser235/236) (Cell Signaling Technology Cat# 2211), Ki-67 (Abcam Cat# ab15580), HER2 (DAKO Cat# A0485), p-HER2 (Y1248) (R&D Systems Cat# AF1768), and p-cMET (Tyr1234/1235) (Cell Signaling Technology Cat# 3129).. Secondary detection was performed with a goat anti-mouse IgG conjugated to DyLight 680 (Thermo Pierce Cat# 35518) and goat anti-rabbit IgG Frontiers in Oncology | www.frontiersin.org 3 conjugated to DyLight 800 (Thermo Pierce Cat# 35571) antibodies.Secondary detection was performed with a goat anti-mouse IgG conjugated to DyLight 680 (Thermo Pierce Cat# 35518) and goat anti-rabbit IgG Frontiers in Oncology | www.frontiersin.org 3 conjugated to DyLight 800 (Thermo Pierce Cat#..

    Expressing:

    Article Title: Ex Vivo Drug Screening Informed Targeted Therapy for Metastatic Parotid Squamous Cell Carcinoma
    Article Snippet: To generate reverse-phase arrays, lysates were printed on nitrocellulose-coated glass slides (Grace Biolabs #305177) on a Genetix QArray Mini Arrayer (Molecular Devices).To generate reverse-phase arrays, lysates were printed on nitrocellulose-coated glass slides (Grace Biolabs #305177) on a Genetix QArray Mini Arrayer (Molecular Devices).. Primary antibodies used for RPPA experiments contained the following: p-AKT (Ser473) (Cell Signaling Technology Cat# 4060), AKT (Cell Signaling Technology Cat# 4691), p-ERK1/2 (Thr202/Tyr204) (Cell Signaling Technology Cat# 4370), p-S6 Ribosomal protein (Ser235/236) (Cell Signaling Technology Cat# 2211), Ki-67 (Abcam Cat# ab15580), HER2 (DAKO Cat# A0485), p-HER2 (Y1248) (R&D Systems Cat# AF1768), and p-cMET (Tyr1234/1235) (Cell Signaling Technology Cat# 3129).. Secondary detection was performed with a goat anti-mouse IgG conjugated to DyLight 680 (Thermo Pierce Cat# 35518) and goat anti-rabbit IgG conjugated to DyLight 800 (Thermo Pierce Cat# 35571) antibodies.Secondary detection was performed with a goat anti-mouse IgG conjugated to DyLight 680 (Thermo Pierce Cat# 35518) and goat anti-rabbit IgG conjugated to DyLight 800 (Thermo Pierce Cat# 35571) antibodies.

    Article Title: Ex Vivo Drug Screening Informed Targeted Therapy for Metastatic Parotid Squamous Cell Carcinoma.
    Article Snippet: To generate reverse-phase arrays, lysates were printed on nitrocellulose-coated glass slides (Grace Biolabs #305177) on a Genetix QArray Mini Arrayer (Molecular Devices).To generate reverse-phase arrays, lysates were printed on nitrocellulose-coated glass slides (Grace Biolabs #305177) on a Genetix QArray Mini Arrayer (Molecular Devices).. Primary antibodies used for RPPA experiments contained the following: p-AKT (Ser473) (Cell Signaling Technology Cat# 4060), AKT (Cell Signaling Technology Cat# 4691), p-ERK1/2 (Thr202/Tyr204) (Cell Signaling Technology Cat# 4370), p-S6 Ribosomal protein (Ser235/236) (Cell Signaling Technology Cat# 2211), Ki-67 (Abcam Cat# ab15580), HER2 (DAKO Cat# A0485), p-HER2 (Y1248) (R&D Systems Cat# AF1768), and p-cMET (Tyr1234/1235) (Cell Signaling Technology Cat# 3129).. Secondary detection was performed with a goat anti-mouse IgG conjugated to DyLight 680 (Thermo Pierce Cat# 35518) and goat anti-rabbit IgG Frontiers in Oncology | www.frontiersin.org 3 conjugated to DyLight 800 (Thermo Pierce Cat# 35571) antibodies.Secondary detection was performed with a goat anti-mouse IgG conjugated to DyLight 680 (Thermo Pierce Cat# 35518) and goat anti-rabbit IgG Frontiers in Oncology | www.frontiersin.org 3 conjugated to DyLight 800 (Thermo Pierce Cat#..



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    R&D Systems p her2 y1248
    Validation of <t>HER2</t> status and targeted therapy. (A) Immunohistochemistry validation of HER2 staining in the affected lymph node. Scale bars: 2.5 mm, 250 µm, and 100 µm (left to right). (B) Radiologic response of patient. Left, prior to treatment, and right, after four cycles of treatment with T-DM1.
    P Her2 Y1248, supplied by R&D Systems, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    A combination of copanlisib and afatinib effectively blocked the phosphorylation of HER2, HER3 and Akt, and enhanced caspase-3 cleavage. (A) Cal27 and (B) FaDu cells were treated with increasing concentrations of copanlisib, 0.5 µM afatinib or their combination for 24 h before lysis. The indicated proteins were detected by western blot analysis. C-, cleaved; P-, phosphorylated.

    Journal: Oncology Reports

    Article Title: Evaluation of co‑inhibition of ErbB family kinases and PI3K for HPV‑negative head and neck squamous cell carcinoma

    doi: 10.3892/or.2025.8871

    Figure Lengend Snippet: A combination of copanlisib and afatinib effectively blocked the phosphorylation of HER2, HER3 and Akt, and enhanced caspase-3 cleavage. (A) Cal27 and (B) FaDu cells were treated with increasing concentrations of copanlisib, 0.5 µM afatinib or their combination for 24 h before lysis. The indicated proteins were detected by western blot analysis. C-, cleaved; P-, phosphorylated.

    Article Snippet: The following antibodies were purchased from Cell Signaling Technology, Inc.: Phosphorylated (P)-Akt-S473 (cat. no. 4058), P-Akt-T308 (cat. no. 9275), Akt (cat. no. 2938), P-HER2-Y1248 (cat. no. 2247), HER2 (cat. no. 4290), P-HER3-Y1289 (cat. no. 2842), HER3 (cat. no. 12708), caspase-3 (cat. no. 9665), cleaved-caspase-3 (cat. no. 9664) and β-actin (cat. no. 4967).

    Techniques: Phospho-proteomics, Lysis, Western Blot

    Structure and function of the HER2 receptor and its inhibition by trastuzumab. ( A – C) show the structure of the receptor, its activation, internalization, recycling and shedding; ( D ) illustrates how the monoclonal antibody trastuzumab (TZ) interacts with HER2 at a site in the IV extracellular domain of the receptor, impairing its function.

    Journal: International Journal of Nanomedicine

    Article Title: Development of High-Loading Trastuzumab PLGA Nanoparticles: A Powerful Tool Against HER2 Positive Breast Cancer Cells

    doi: 10.2147/IJN.S429898

    Figure Lengend Snippet: Structure and function of the HER2 receptor and its inhibition by trastuzumab. ( A – C) show the structure of the receptor, its activation, internalization, recycling and shedding; ( D ) illustrates how the monoclonal antibody trastuzumab (TZ) interacts with HER2 at a site in the IV extracellular domain of the receptor, impairing its function.

    Article Snippet: Antibodies to p-HER2 (Y1248 # 2247), HER2 N-term (#4290) HER2 (#3250), p-AKT (S473) (#9271), AKT (#9272) were from Cell Signaling Technology (Beverly, MA USA); β-Actin (F-3022) and α-Tubulin (T5168) were from Sigma-Aldrich Co. (Merck KGaA, St. Louis, MO, USA).

    Techniques: Inhibition, Activation Assay

    Biological effects of encapsulated or free trastuzumab on SKBR3 cells. ( A ) Western blot assessment of HER2 Y1248 and AKT S473 phosphorylation on extracts from SKBR3 cells exposed to NP or two increasing concentrations of TZPs or free TZ for the indicated time points. The histograms report the quantification of the phosphorylation levels normalized to total HER2 or AKT, respectively; FC indicates Fold-Change. The tables below illustrate the results obtained by comparing the single dose of each formulation to all others using the same colour code as in the panel on the right side of the figure. ( B ) Viability of SKBR3 cells treated with NPs or two increasing concentrations of TZPs or free TZ for the indicated time points reported as percentage vs untreated CTR control cells. Data are reported as mean± SEM of two independent experiments performed at least in triplicate; the mean values from untreated or treated cells were compared by applying the t -test. Also, in this case, the tables below illustrate the results obtained by comparing the single dose of each formulation to all others using the same colour code as in the panel on the right side of the figure. Statistical significance was considered when * p <0.05, ** p < 0.01, ***p < 0.001. **** p < 0.0001, ns= not significant.

    Journal: International Journal of Nanomedicine

    Article Title: Development of High-Loading Trastuzumab PLGA Nanoparticles: A Powerful Tool Against HER2 Positive Breast Cancer Cells

    doi: 10.2147/IJN.S429898

    Figure Lengend Snippet: Biological effects of encapsulated or free trastuzumab on SKBR3 cells. ( A ) Western blot assessment of HER2 Y1248 and AKT S473 phosphorylation on extracts from SKBR3 cells exposed to NP or two increasing concentrations of TZPs or free TZ for the indicated time points. The histograms report the quantification of the phosphorylation levels normalized to total HER2 or AKT, respectively; FC indicates Fold-Change. The tables below illustrate the results obtained by comparing the single dose of each formulation to all others using the same colour code as in the panel on the right side of the figure. ( B ) Viability of SKBR3 cells treated with NPs or two increasing concentrations of TZPs or free TZ for the indicated time points reported as percentage vs untreated CTR control cells. Data are reported as mean± SEM of two independent experiments performed at least in triplicate; the mean values from untreated or treated cells were compared by applying the t -test. Also, in this case, the tables below illustrate the results obtained by comparing the single dose of each formulation to all others using the same colour code as in the panel on the right side of the figure. Statistical significance was considered when * p <0.05, ** p < 0.01, ***p < 0.001. **** p < 0.0001, ns= not significant.

    Article Snippet: Antibodies to p-HER2 (Y1248 # 2247), HER2 N-term (#4290) HER2 (#3250), p-AKT (S473) (#9271), AKT (#9272) were from Cell Signaling Technology (Beverly, MA USA); β-Actin (F-3022) and α-Tubulin (T5168) were from Sigma-Aldrich Co. (Merck KGaA, St. Louis, MO, USA).

    Techniques: Western Blot, Formulation

    TZPs and TZ promote HER2 endocytosis and reduce shedding. ( A ) Flow cytometry analysis of cell surface HER2 in SKBR3 cells untreated (black line) or treated with NPs (grey line) or two increasing concentrations of TZPs (heavy and light green line) or free TZ (heavy and light orange line) for 24 h. The percentage of cell counts are reported on the y axis, while the Fluorescence Intensity on the x axis. The right-most panel is the merge of the single panels. ( B ) The histograms report the quantification of HER2 levels, expressed as percentage vs untreated controls, after treatment with NPs (grey), TZPs (heavy and light green) or free TZ (heavy and light orange). The tables below illustrate the results obtained by comparing the single dose of each formulation to all others, using the same colour code as in the panel on the right side of the figure. Statistical significance was considered when * p < 0.05, ** p < 0.01, *** p < 0.001. **** p < 0.0001, ns= not significant. ( C ) Western blot analysis of total HER2 protein in SKBR3 cells untreated or treated as above for the indicated time points. The histograms report the quantification after normalization to α-Tubulin used as protein loading control. The tables below illustrate the results obtained by comparing the single dose of each formulation to all others, using the same colour code as in the panel on the right side of the figure. Statistical significance was considered when * p < 0.05, ** p < 0.01, *** p < 0.001. **** p < 0.0001, ns= not significant. ( D ) Western blot analysis of the soluble form of HER2 (sHER2) present in the concentrated conditioned media of SKBR3 cells treated with NPs or TZPs as above for the indicated time points. The histograms show the relative quantification after normalization to protein loading through Ponceau S staining of the filter. The tables below illustrate the results obtained by comparing the single dose of each formulation to all others, using the same colour code as in the panel on the right side of the figure. Statistical significance was considered when * p < 0.05, ** p < 0.01, *** p < 0.001. **** p < 0.0001, ns= not significant.

    Journal: International Journal of Nanomedicine

    Article Title: Development of High-Loading Trastuzumab PLGA Nanoparticles: A Powerful Tool Against HER2 Positive Breast Cancer Cells

    doi: 10.2147/IJN.S429898

    Figure Lengend Snippet: TZPs and TZ promote HER2 endocytosis and reduce shedding. ( A ) Flow cytometry analysis of cell surface HER2 in SKBR3 cells untreated (black line) or treated with NPs (grey line) or two increasing concentrations of TZPs (heavy and light green line) or free TZ (heavy and light orange line) for 24 h. The percentage of cell counts are reported on the y axis, while the Fluorescence Intensity on the x axis. The right-most panel is the merge of the single panels. ( B ) The histograms report the quantification of HER2 levels, expressed as percentage vs untreated controls, after treatment with NPs (grey), TZPs (heavy and light green) or free TZ (heavy and light orange). The tables below illustrate the results obtained by comparing the single dose of each formulation to all others, using the same colour code as in the panel on the right side of the figure. Statistical significance was considered when * p < 0.05, ** p < 0.01, *** p < 0.001. **** p < 0.0001, ns= not significant. ( C ) Western blot analysis of total HER2 protein in SKBR3 cells untreated or treated as above for the indicated time points. The histograms report the quantification after normalization to α-Tubulin used as protein loading control. The tables below illustrate the results obtained by comparing the single dose of each formulation to all others, using the same colour code as in the panel on the right side of the figure. Statistical significance was considered when * p < 0.05, ** p < 0.01, *** p < 0.001. **** p < 0.0001, ns= not significant. ( D ) Western blot analysis of the soluble form of HER2 (sHER2) present in the concentrated conditioned media of SKBR3 cells treated with NPs or TZPs as above for the indicated time points. The histograms show the relative quantification after normalization to protein loading through Ponceau S staining of the filter. The tables below illustrate the results obtained by comparing the single dose of each formulation to all others, using the same colour code as in the panel on the right side of the figure. Statistical significance was considered when * p < 0.05, ** p < 0.01, *** p < 0.001. **** p < 0.0001, ns= not significant.

    Article Snippet: Antibodies to p-HER2 (Y1248 # 2247), HER2 N-term (#4290) HER2 (#3250), p-AKT (S473) (#9271), AKT (#9272) were from Cell Signaling Technology (Beverly, MA USA); β-Actin (F-3022) and α-Tubulin (T5168) were from Sigma-Aldrich Co. (Merck KGaA, St. Louis, MO, USA).

    Techniques: Flow Cytometry, Fluorescence, Formulation, Western Blot, Staining

    KEY RESOURCES TABLE

    Journal: Cell reports

    Article Title: Extensive conformational and physical plasticity protects HER2-HER3 tumorigenic signaling

    doi: 10.1016/j.celrep.2021.110285

    Figure Lengend Snippet: KEY RESOURCES TABLE

    Article Snippet: anti-p-HER2 Y1248 , Cell Signaling Technology , 2247.

    Techniques: Recombinant, Transfection, Plasmid Preparation, BIA-KA, Mutagenesis, Modification

    Validation of HER2 status and targeted therapy. (A) Immunohistochemistry validation of HER2 staining in the affected lymph node. Scale bars: 2.5 mm, 250 µm, and 100 µm (left to right). (B) Radiologic response of patient. Left, prior to treatment, and right, after four cycles of treatment with T-DM1.

    Journal: Frontiers in Oncology

    Article Title: Ex Vivo Drug Screening Informed Targeted Therapy for Metastatic Parotid Squamous Cell Carcinoma

    doi: 10.3389/fonc.2021.735820

    Figure Lengend Snippet: Validation of HER2 status and targeted therapy. (A) Immunohistochemistry validation of HER2 staining in the affected lymph node. Scale bars: 2.5 mm, 250 µm, and 100 µm (left to right). (B) Radiologic response of patient. Left, prior to treatment, and right, after four cycles of treatment with T-DM1.

    Article Snippet: Primary antibodies used for RPPA experiments contained the following: p-AKT (Ser473) (Cell Signaling Technology Cat# 4060), AKT (Cell Signaling Technology Cat# 4691), p-ERK1/2 (Thr202/Tyr204) (Cell Signaling Technology Cat# 4370), p-S6 Ribosomal protein (Ser235/236) (Cell Signaling Technology Cat# 2211), Ki-67 (Abcam Cat# ab15580), HER2 (DAKO Cat# A0485), p-HER2 (Y1248) (R&D Systems Cat# AF1768), and p-cMET (Tyr1234/1235) (Cell Signaling Technology Cat# 3129).

    Techniques: Biomarker Discovery, Immunohistochemistry, Staining

    Repeat ex vivo drug screen in recurrent disease. (A) Scatter plot showing correlation of the mean GR score of 193 drugs in five doses for the HER2 + and VIM + cells. (B) Scatter plot showing correlation of the IC 50 estimate of the analyzed drugs between the HER2 + sample B cells and KRT19 + sample A cells. (C) Box plots showing the GR scores for the five doses of afatinib, neratinib, erlotinib, and lapatinib in sample A and B KRT19 + and HER + cells. Afatinib and neratinib displayed a statistically significant differential effect on the KRT19 + and HER + cells from sample B as compared using paired Student’s t -test. (D) Gallery view of representative 20× immunofluorescence microscopy images of the sample cells treated for 96 h with the different EGFR-TKis in five different concentrations (nM). KRT19 staining shown in blue, VIM staining in green and HER2 staining in red. Bars: 50 µm.

    Journal: Frontiers in Oncology

    Article Title: Ex Vivo Drug Screening Informed Targeted Therapy for Metastatic Parotid Squamous Cell Carcinoma

    doi: 10.3389/fonc.2021.735820

    Figure Lengend Snippet: Repeat ex vivo drug screen in recurrent disease. (A) Scatter plot showing correlation of the mean GR score of 193 drugs in five doses for the HER2 + and VIM + cells. (B) Scatter plot showing correlation of the IC 50 estimate of the analyzed drugs between the HER2 + sample B cells and KRT19 + sample A cells. (C) Box plots showing the GR scores for the five doses of afatinib, neratinib, erlotinib, and lapatinib in sample A and B KRT19 + and HER + cells. Afatinib and neratinib displayed a statistically significant differential effect on the KRT19 + and HER + cells from sample B as compared using paired Student’s t -test. (D) Gallery view of representative 20× immunofluorescence microscopy images of the sample cells treated for 96 h with the different EGFR-TKis in five different concentrations (nM). KRT19 staining shown in blue, VIM staining in green and HER2 staining in red. Bars: 50 µm.

    Article Snippet: Primary antibodies used for RPPA experiments contained the following: p-AKT (Ser473) (Cell Signaling Technology Cat# 4060), AKT (Cell Signaling Technology Cat# 4691), p-ERK1/2 (Thr202/Tyr204) (Cell Signaling Technology Cat# 4370), p-S6 Ribosomal protein (Ser235/236) (Cell Signaling Technology Cat# 2211), Ki-67 (Abcam Cat# ab15580), HER2 (DAKO Cat# A0485), p-HER2 (Y1248) (R&D Systems Cat# AF1768), and p-cMET (Tyr1234/1235) (Cell Signaling Technology Cat# 3129).

    Techniques: Ex Vivo, Immunofluorescence, Microscopy, Staining

    A pathway perspective on targeted therapy responses. (A) Heat map visualization with one-dimensional (vertical) unsupervised clustering of the z -scores of the eight RPPA markers across 19 drugs in five concentrations (left to right, low to high, respectively). (B) Top: Representative microscope images of HER2 staining on sample B cells in the ex vivo screen following 96-h exposure to the indicated drugs at 5 µM concentration. KRT19 staining shown in blue, VIM staining in green, and HER2 staining in red. Bars: 50 µm. Bottom left: Box plot showing the mean and standard deviation of fluorescence intensity of HER2 staining (RFU) of the KRT19 + cells in response to treatment with the different EGFR-TKis. Two biological replicate experiments were combined and compared using unpaired Welch’s t -test. Bottom right: Comparison of the HER2 staining intensity to DMSO-treated cells. Signal shown as fold change. (C) Combination of lapatinib and afatanib shows a synergistic growth inhibitory effect. With all three different drug schedules tested, the combination resulted in significantly increased efficacy over the single agents as indicated by the lower IC 50 (middle panel). On the basis of the CI 50 combination index, lapatinib pre-treatment preceding afatinib treatment had the highest synergy (right panel). Comparison was made using unpaired Student’s t -test. (D) The patient-derived tumor cells were treated with DMSO (−), 1000 nmol/L afatinib, lapatinib, neratinib, or combination of 1000 nmol/L afatinib+lapatinib, or neratinib+lapatinib for 48 h, and phosphorylation (p) AKT and total HER2 protein levels of indicated markers were assessed using Western blot. Actin was assessed as a loading control.

    Journal: Frontiers in Oncology

    Article Title: Ex Vivo Drug Screening Informed Targeted Therapy for Metastatic Parotid Squamous Cell Carcinoma

    doi: 10.3389/fonc.2021.735820

    Figure Lengend Snippet: A pathway perspective on targeted therapy responses. (A) Heat map visualization with one-dimensional (vertical) unsupervised clustering of the z -scores of the eight RPPA markers across 19 drugs in five concentrations (left to right, low to high, respectively). (B) Top: Representative microscope images of HER2 staining on sample B cells in the ex vivo screen following 96-h exposure to the indicated drugs at 5 µM concentration. KRT19 staining shown in blue, VIM staining in green, and HER2 staining in red. Bars: 50 µm. Bottom left: Box plot showing the mean and standard deviation of fluorescence intensity of HER2 staining (RFU) of the KRT19 + cells in response to treatment with the different EGFR-TKis. Two biological replicate experiments were combined and compared using unpaired Welch’s t -test. Bottom right: Comparison of the HER2 staining intensity to DMSO-treated cells. Signal shown as fold change. (C) Combination of lapatinib and afatanib shows a synergistic growth inhibitory effect. With all three different drug schedules tested, the combination resulted in significantly increased efficacy over the single agents as indicated by the lower IC 50 (middle panel). On the basis of the CI 50 combination index, lapatinib pre-treatment preceding afatinib treatment had the highest synergy (right panel). Comparison was made using unpaired Student’s t -test. (D) The patient-derived tumor cells were treated with DMSO (−), 1000 nmol/L afatinib, lapatinib, neratinib, or combination of 1000 nmol/L afatinib+lapatinib, or neratinib+lapatinib for 48 h, and phosphorylation (p) AKT and total HER2 protein levels of indicated markers were assessed using Western blot. Actin was assessed as a loading control.

    Article Snippet: Primary antibodies used for RPPA experiments contained the following: p-AKT (Ser473) (Cell Signaling Technology Cat# 4060), AKT (Cell Signaling Technology Cat# 4691), p-ERK1/2 (Thr202/Tyr204) (Cell Signaling Technology Cat# 4370), p-S6 Ribosomal protein (Ser235/236) (Cell Signaling Technology Cat# 2211), Ki-67 (Abcam Cat# ab15580), HER2 (DAKO Cat# A0485), p-HER2 (Y1248) (R&D Systems Cat# AF1768), and p-cMET (Tyr1234/1235) (Cell Signaling Technology Cat# 3129).

    Techniques: Microscopy, Staining, Ex Vivo, Concentration Assay, Standard Deviation, Fluorescence, Comparison, Derivative Assay, Phospho-proteomics, Western Blot, Control

    Flow cytometry immunophenotyping of the MISB10B cells. (A) Analysis of expression of epithelial lineage markers and HER2 in the EpCAM + cells. (B) Representative bright-field microscopy image (10×) of the morphology of the MISB10B cells in near full confluent state. (C) Analysis of expression of additional immunophenotypic markers in the EpCAM + cells.

    Journal: Frontiers in Oncology

    Article Title: Ex Vivo Drug Screening Informed Targeted Therapy for Metastatic Parotid Squamous Cell Carcinoma

    doi: 10.3389/fonc.2021.735820

    Figure Lengend Snippet: Flow cytometry immunophenotyping of the MISB10B cells. (A) Analysis of expression of epithelial lineage markers and HER2 in the EpCAM + cells. (B) Representative bright-field microscopy image (10×) of the morphology of the MISB10B cells in near full confluent state. (C) Analysis of expression of additional immunophenotypic markers in the EpCAM + cells.

    Article Snippet: Primary antibodies used for RPPA experiments contained the following: p-AKT (Ser473) (Cell Signaling Technology Cat# 4060), AKT (Cell Signaling Technology Cat# 4691), p-ERK1/2 (Thr202/Tyr204) (Cell Signaling Technology Cat# 4370), p-S6 Ribosomal protein (Ser235/236) (Cell Signaling Technology Cat# 2211), Ki-67 (Abcam Cat# ab15580), HER2 (DAKO Cat# A0485), p-HER2 (Y1248) (R&D Systems Cat# AF1768), and p-cMET (Tyr1234/1235) (Cell Signaling Technology Cat# 3129).

    Techniques: Flow Cytometry, Expressing, Microscopy