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rabbit anti phospho erbb4 (Bioss)

Name: rabbit anti phospho erbb4
Brand: Bioss
Rating: 94 (4 reviews)

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Bioss rabbit anti phospho erbb4

The effects of <t>ErbB4</t> receptor agonist E4A and melatonin on cognitive and spatial memory deficits in D-gal-induced aging mice. (A) Representative swimming trajectories of mice subjected to different treatments in the Morris Water Maze (MWM) spatial probe test. (B) Escape latency across the five experimental groups. (C) Number of target crossings during the MWM probe test. (D) Time spent in the target quadrant during the MWM probe test. (E) Representative heat map showing interaction frequency with novel versus familiar objects. (F) Recognition index in the Novel Object Recognition test among the experimental groups. (G) Spontaneous alternation behaviors in the Y-maze test were assessed. Data were presented as mean ± SD (n = 6–8). Statistical significance was denoted as follows: *p < 0.05, **p < 0.01, ****p < 0.0001.

Rabbit Anti Phospho Erbb4, supplied by Bioss, used in various techniques. Bioz Stars score: 94/100, based on 4 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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Average 94 stars, based on 4 article reviews

rabbit anti phospho erbb4 - by Bioz Stars, 2026-02

94/100 stars

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1) Product Images from "Targeted ErbB4 receptor activation prevents D-galactose-induced neuronal senescence via inhibiting ferroptosis pathway"

Article Title: Targeted ErbB4 receptor activation prevents D-galactose-induced neuronal senescence via inhibiting ferroptosis pathway

Journal: Frontiers in Pharmacology

doi: 10.3389/fphar.2025.1528604

The effects of ErbB4 receptor agonist E4A and melatonin on cognitive and spatial memory deficits in D-gal-induced aging mice. (A) Representative swimming trajectories of mice subjected to different treatments in the Morris Water Maze (MWM) spatial probe test. (B) Escape latency across the five experimental groups. (C) Number of target crossings during the MWM probe test. (D) Time spent in the target quadrant during the MWM probe test. (E) Representative heat map showing interaction frequency with novel versus familiar objects. (F) Recognition index in the Novel Object Recognition test among the experimental groups. (G) Spontaneous alternation behaviors in the Y-maze test were assessed. Data were presented as mean ± SD (n = 6–8). Statistical significance was denoted as follows: *p < 0.05, **p < 0.01, ****p < 0.0001.

Figure Legend Snippet: The effects of ErbB4 receptor agonist E4A and melatonin on cognitive and spatial memory deficits in D-gal-induced aging mice. (A) Representative swimming trajectories of mice subjected to different treatments in the Morris Water Maze (MWM) spatial probe test. (B) Escape latency across the five experimental groups. (C) Number of target crossings during the MWM probe test. (D) Time spent in the target quadrant during the MWM probe test. (E) Representative heat map showing interaction frequency with novel versus familiar objects. (F) Recognition index in the Novel Object Recognition test among the experimental groups. (G) Spontaneous alternation behaviors in the Y-maze test were assessed. Data were presented as mean ± SD (n = 6–8). Statistical significance was denoted as follows: *p < 0.05, **p < 0.01, ****p < 0.0001.

Techniques Used:

Effect of ErbB4 receptor agonist and melatonin can ameliorate D-gal-induced aging in hippocampus in mice. (A) Western blot analysis of Lamin B1, p53, p21, p16, GFAP, and Iba-1 protein expression levels in the hippocampus of mice. (B–G) Quantification of Lamin B1, p53, p21, p16, GFAP, and Iba-1 protein levels. Data are presented as mean ± SD, with n = 4–5. *p < 0.05, **p < 0.01, ***p < 0.001.

Figure Legend Snippet: Effect of ErbB4 receptor agonist and melatonin can ameliorate D-gal-induced aging in hippocampus in mice. (A) Western blot analysis of Lamin B1, p53, p21, p16, GFAP, and Iba-1 protein expression levels in the hippocampus of mice. (B–G) Quantification of Lamin B1, p53, p21, p16, GFAP, and Iba-1 protein levels. Data are presented as mean ± SD, with n = 4–5. *p < 0.05, **p < 0.01, ***p < 0.001.

Techniques Used: Western Blot, Expressing

ErbB4 receptor agonist and melatonin inhibit ferroptosis in hippocampus in D-gal-induced aging mice. (A) Immunofluorescence analysis of GPX4-positive cells, SLC7A11-positive cells, TFRC-positive cells in the dentate gyrus (DG) and CA1 regions. Scale bar = 50 μm. (B) Western blot analysis of Nrf2, TFRC, SLC7A11, and GPX4 protein expression levels in the hippocampus of mice. (C–F) Quantification of Nrf2, TFRC, SLC7A11, and GPX4 protein levels. Data are presented as mean ± SD, with n = 4–5. *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001.

Figure Legend Snippet: ErbB4 receptor agonist and melatonin inhibit ferroptosis in hippocampus in D-gal-induced aging mice. (A) Immunofluorescence analysis of GPX4-positive cells, SLC7A11-positive cells, TFRC-positive cells in the dentate gyrus (DG) and CA1 regions. Scale bar = 50 μm. (B) Western blot analysis of Nrf2, TFRC, SLC7A11, and GPX4 protein expression levels in the hippocampus of mice. (C–F) Quantification of Nrf2, TFRC, SLC7A11, and GPX4 protein levels. Data are presented as mean ± SD, with n = 4–5. *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001.

Techniques Used: Immunofluorescence, Western Blot, Expressing

The mitigation of D-gal-induced ferroptosis in HT22 cells by ErbB4 receptor agonist and melatonin. (A) Cell viability post D-gal exposure, evaluated using the CCK-8 assay (n = 5). (B) Cell viability following co-treatment with D-gal and ErbB4 receptor agonist, assessed via the CCK-8 assay (n = 6). (C) Cell viability following co-treatment with D-gal and melatonin, determined by the CCK-8 assay (n = 6). (D) Visualization of cellular senescence through SA-β-gal staining. Scale bar = 100 μm. (E) Quantification of SA-β-gal staining intensities (n = 3). (F) Western blot analysis was conducted to assess the expression levels of senescence-associated markers and ferroptosis-related proteins in HT22 cells. (G–M) Quantification of Nrf2, TFRC, SLC7A11, GPX4, p53, p21, and p16 levels was performed, with normalization to GAPDH (n = 4–7). (N) Intracellular GSH levels (n = 4). (O) Intracellular MDA levels (n = 5). (P) Quantitative analysis of ROS levels was undertaken (n = 4). (Q) Intracellular ROS generation was measured using DCFH-DA, wherein ROS activity was reflected as green fluorescence intensity. Scale bar = 200 μm. Data are presented as mean ± SD. Statistical significance is indicated as follows: *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001.

Figure Legend Snippet: The mitigation of D-gal-induced ferroptosis in HT22 cells by ErbB4 receptor agonist and melatonin. (A) Cell viability post D-gal exposure, evaluated using the CCK-8 assay (n = 5). (B) Cell viability following co-treatment with D-gal and ErbB4 receptor agonist, assessed via the CCK-8 assay (n = 6). (C) Cell viability following co-treatment with D-gal and melatonin, determined by the CCK-8 assay (n = 6). (D) Visualization of cellular senescence through SA-β-gal staining. Scale bar = 100 μm. (E) Quantification of SA-β-gal staining intensities (n = 3). (F) Western blot analysis was conducted to assess the expression levels of senescence-associated markers and ferroptosis-related proteins in HT22 cells. (G–M) Quantification of Nrf2, TFRC, SLC7A11, GPX4, p53, p21, and p16 levels was performed, with normalization to GAPDH (n = 4–7). (N) Intracellular GSH levels (n = 4). (O) Intracellular MDA levels (n = 5). (P) Quantitative analysis of ROS levels was undertaken (n = 4). (Q) Intracellular ROS generation was measured using DCFH-DA, wherein ROS activity was reflected as green fluorescence intensity. Scale bar = 200 μm. Data are presented as mean ± SD. Statistical significance is indicated as follows: *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001.

Techniques Used: CCK-8 Assay, Staining, Western Blot, Expressing, Activity Assay, Fluorescence

The effects of ErbB4 receptor agonist and melatonin treatment on ferroptosis in Erastin-exposed HT22 cells. (A) Cell viability of HT22 cells post-Erastin exposure was evaluated using the CCK-8 assay (n = 5). (B) Cell viability was evaluated following treatment with Erastin, cotreatment with either E4A or melatonin, or both, utilizing the CCK-8 assay (n = 4). (C) Western blot analysis was conducted to determine the expression levels of senescence-associated markers and ferroptosis-related proteins in HT22 cells. (D–I) Quantitative analysis of Nrf2, TFRC, SLC7A11, GPX4, p21, and p16 levels, normalized to GAPDH (n = 4–7). (J) Intracellular ROS generation was measured using DCFH-DA, with ROS activity indicated by green fluorescence. Scale bar = 200 μm. (K) The quantitative analysis of ROS levels was conducted (n = 4). The data are presented as mean ± SD. Statistical significance is indicated as follows: *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001.

Figure Legend Snippet: The effects of ErbB4 receptor agonist and melatonin treatment on ferroptosis in Erastin-exposed HT22 cells. (A) Cell viability of HT22 cells post-Erastin exposure was evaluated using the CCK-8 assay (n = 5). (B) Cell viability was evaluated following treatment with Erastin, cotreatment with either E4A or melatonin, or both, utilizing the CCK-8 assay (n = 4). (C) Western blot analysis was conducted to determine the expression levels of senescence-associated markers and ferroptosis-related proteins in HT22 cells. (D–I) Quantitative analysis of Nrf2, TFRC, SLC7A11, GPX4, p21, and p16 levels, normalized to GAPDH (n = 4–7). (J) Intracellular ROS generation was measured using DCFH-DA, with ROS activity indicated by green fluorescence. Scale bar = 200 μm. (K) The quantitative analysis of ROS levels was conducted (n = 4). The data are presented as mean ± SD. Statistical significance is indicated as follows: *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001.

Techniques Used: CCK-8 Assay, Western Blot, Expressing, Activity Assay, Fluorescence

ErbB4 receptor agonist mitigates D-gal-induced hippocampal neuronal aging both in vivo and in vitro through the activation of ErbB4 receptors and modulation of the Akt/Nrf2 signaling pathway. (A) Western blot analysis of pErbB4, ErbB4, pAkt1, and Akt1 protein expression levels in the hippocampus of mice. (B, C) Quantification of pErbB4 and pAkt1 levels (n = 4). (D) Western blot analysis of pErbB4, ErbB4, pAkt1, and Akt1 protein expression levels in D-gal-induced HT22 cells. (E, F) Quantification of pErbB4 and pAkt1 levels (n = 4). (G, H) Immunofluorescence analysis of the Nrf2-positive cells in the DG region (G) and in the cortex region (H) . Scale bar = 50 μm. (I) Western blot analysis of Nrf2, SLC7A11, GPX4, Lamin B1 and P21 protein levels after treatment of Nrf2 inhibitor AEM1. (J–N) Quantitative analysis of Nrf2, SLC7A11, GPX4, Lamin B1 and P21 levels (n = 4–7). Data are presented as mean ± SD. Statistical significance is indicated as follows: *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001.

Figure Legend Snippet: ErbB4 receptor agonist mitigates D-gal-induced hippocampal neuronal aging both in vivo and in vitro through the activation of ErbB4 receptors and modulation of the Akt/Nrf2 signaling pathway. (A) Western blot analysis of pErbB4, ErbB4, pAkt1, and Akt1 protein expression levels in the hippocampus of mice. (B, C) Quantification of pErbB4 and pAkt1 levels (n = 4). (D) Western blot analysis of pErbB4, ErbB4, pAkt1, and Akt1 protein expression levels in D-gal-induced HT22 cells. (E, F) Quantification of pErbB4 and pAkt1 levels (n = 4). (G, H) Immunofluorescence analysis of the Nrf2-positive cells in the DG region (G) and in the cortex region (H) . Scale bar = 50 μm. (I) Western blot analysis of Nrf2, SLC7A11, GPX4, Lamin B1 and P21 protein levels after treatment of Nrf2 inhibitor AEM1. (J–N) Quantitative analysis of Nrf2, SLC7A11, GPX4, Lamin B1 and P21 levels (n = 4–7). Data are presented as mean ± SD. Statistical significance is indicated as follows: *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001.

Techniques Used: In Vivo, In Vitro, Activation Assay, Western Blot, Expressing, Immunofluorescence

Schematic diagram illustrating the effects of targeted activation of ErbB4 receptor on neuronal aging via ferroptosis inhibition. Small molecule agonist (E4A)can activate ErbB4 receptors and regulate the Akt/Nrf2 signaling pathway to achieve neuroprotective effect in D-gal-induced neuronal aging. D-gal treatment increases the accumulation of advanced glycation end products (AGEs) which stimulates the production of reactive oxygen species (ROS) and increase the expression of senescence marker genes P53, P16, and P21. E4A can promote Akt1 phosphorylation and promote Nrf2 entrance into the nucleus, in which it involves in regulating the expression of ferroptosis suppressor gene, such as SLC7A11 and GPX4 to attenuate lipid peroxidation, which can inhibit cell ferroptosis and neuronal aging. The diagram was created with MedPeer ( www.medpeer.cn ).

Figure Legend Snippet: Schematic diagram illustrating the effects of targeted activation of ErbB4 receptor on neuronal aging via ferroptosis inhibition. Small molecule agonist (E4A)can activate ErbB4 receptors and regulate the Akt/Nrf2 signaling pathway to achieve neuroprotective effect in D-gal-induced neuronal aging. D-gal treatment increases the accumulation of advanced glycation end products (AGEs) which stimulates the production of reactive oxygen species (ROS) and increase the expression of senescence marker genes P53, P16, and P21. E4A can promote Akt1 phosphorylation and promote Nrf2 entrance into the nucleus, in which it involves in regulating the expression of ferroptosis suppressor gene, such as SLC7A11 and GPX4 to attenuate lipid peroxidation, which can inhibit cell ferroptosis and neuronal aging. The diagram was created with MedPeer ( www.medpeer.cn ).

Techniques Used: Activation Assay, Inhibition, Expressing, Marker

Image Search Results

Journal: Frontiers in Pharmacology

Article Title: Targeted ErbB4 receptor activation prevents D-galactose-induced neuronal senescence via inhibiting ferroptosis pathway

doi: 10.3389/fphar.2025.1528604

Figure Lengend Snippet: The effects of ErbB4 receptor agonist E4A and melatonin on cognitive and spatial memory deficits in D-gal-induced aging mice. (A) Representative swimming trajectories of mice subjected to different treatments in the Morris Water Maze (MWM) spatial probe test. (B) Escape latency across the five experimental groups. (C) Number of target crossings during the MWM probe test. (D) Time spent in the target quadrant during the MWM probe test. (E) Representative heat map showing interaction frequency with novel versus familiar objects. (F) Recognition index in the Novel Object Recognition test among the experimental groups. (G) Spontaneous alternation behaviors in the Y-maze test were assessed. Data were presented as mean ± SD (n = 6–8). Statistical significance was denoted as follows: *p < 0.05, **p < 0.01, ****p < 0.0001.

Article Snippet: The antibodies used, diluted in primary antibody diluents (P0023A, Beyotime), included: rabbit anti-phospho-ErbB4 (bs-3220R, Bioss; 1:1,000), rabbit anti-ErbB4 (AF6807, Beyotime; 1:1,000), rabbit anti-phospho-Akt1 (AF1546, Beyotime; 1:1,000), mouse anti-Akt1 (sc-377457, Santa Cruz Biotech; 1:1,000), rabbit anti-Nrf2 (AF7623, Beyotime; 1:1,000), rabbit anti-TFRC (AF8136, Beyotime; 1:1,000), rabbit anti-SLC7A11 (GB115276, Servicebio Biotech; 1:1,000), rabbit anti-GPX4 (AF7020, Beyotime; 1:1,000), rabbit anti-Lamin B1 (12987-1-AP, Proteintech, 1:5,000), rabbit anti-p53 (AF5258, Beyotime; 1:1,000), mouse anti-p21 (AP021, Beyotime; 1:1,000), rabbit anti-p16 (bs-23797R, Bioss Biotech, Beijing, China; 1:1,000), and rabbit anti-GAPDH (GB15002, Servicebio Biotech; 1:2000).

Techniques:

Journal: Frontiers in Pharmacology

Article Title: Targeted ErbB4 receptor activation prevents D-galactose-induced neuronal senescence via inhibiting ferroptosis pathway

doi: 10.3389/fphar.2025.1528604

Figure Lengend Snippet: Effect of ErbB4 receptor agonist and melatonin can ameliorate D-gal-induced aging in hippocampus in mice. (A) Western blot analysis of Lamin B1, p53, p21, p16, GFAP, and Iba-1 protein expression levels in the hippocampus of mice. (B–G) Quantification of Lamin B1, p53, p21, p16, GFAP, and Iba-1 protein levels. Data are presented as mean ± SD, with n = 4–5. *p < 0.05, **p < 0.01, ***p < 0.001.

Techniques: Western Blot, Expressing

Journal: Frontiers in Pharmacology

Article Title: Targeted ErbB4 receptor activation prevents D-galactose-induced neuronal senescence via inhibiting ferroptosis pathway

doi: 10.3389/fphar.2025.1528604

Figure Lengend Snippet: ErbB4 receptor agonist and melatonin inhibit ferroptosis in hippocampus in D-gal-induced aging mice. (A) Immunofluorescence analysis of GPX4-positive cells, SLC7A11-positive cells, TFRC-positive cells in the dentate gyrus (DG) and CA1 regions. Scale bar = 50 μm. (B) Western blot analysis of Nrf2, TFRC, SLC7A11, and GPX4 protein expression levels in the hippocampus of mice. (C–F) Quantification of Nrf2, TFRC, SLC7A11, and GPX4 protein levels. Data are presented as mean ± SD, with n = 4–5. *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001.

Techniques: Immunofluorescence, Western Blot, Expressing

Journal: Frontiers in Pharmacology

Article Title: Targeted ErbB4 receptor activation prevents D-galactose-induced neuronal senescence via inhibiting ferroptosis pathway

doi: 10.3389/fphar.2025.1528604

Figure Lengend Snippet: The mitigation of D-gal-induced ferroptosis in HT22 cells by ErbB4 receptor agonist and melatonin. (A) Cell viability post D-gal exposure, evaluated using the CCK-8 assay (n = 5). (B) Cell viability following co-treatment with D-gal and ErbB4 receptor agonist, assessed via the CCK-8 assay (n = 6). (C) Cell viability following co-treatment with D-gal and melatonin, determined by the CCK-8 assay (n = 6). (D) Visualization of cellular senescence through SA-β-gal staining. Scale bar = 100 μm. (E) Quantification of SA-β-gal staining intensities (n = 3). (F) Western blot analysis was conducted to assess the expression levels of senescence-associated markers and ferroptosis-related proteins in HT22 cells. (G–M) Quantification of Nrf2, TFRC, SLC7A11, GPX4, p53, p21, and p16 levels was performed, with normalization to GAPDH (n = 4–7). (N) Intracellular GSH levels (n = 4). (O) Intracellular MDA levels (n = 5). (P) Quantitative analysis of ROS levels was undertaken (n = 4). (Q) Intracellular ROS generation was measured using DCFH-DA, wherein ROS activity was reflected as green fluorescence intensity. Scale bar = 200 μm. Data are presented as mean ± SD. Statistical significance is indicated as follows: *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001.

Techniques: CCK-8 Assay, Staining, Western Blot, Expressing, Activity Assay, Fluorescence

Journal: Frontiers in Pharmacology

Article Title: Targeted ErbB4 receptor activation prevents D-galactose-induced neuronal senescence via inhibiting ferroptosis pathway

doi: 10.3389/fphar.2025.1528604

Figure Lengend Snippet: The effects of ErbB4 receptor agonist and melatonin treatment on ferroptosis in Erastin-exposed HT22 cells. (A) Cell viability of HT22 cells post-Erastin exposure was evaluated using the CCK-8 assay (n = 5). (B) Cell viability was evaluated following treatment with Erastin, cotreatment with either E4A or melatonin, or both, utilizing the CCK-8 assay (n = 4). (C) Western blot analysis was conducted to determine the expression levels of senescence-associated markers and ferroptosis-related proteins in HT22 cells. (D–I) Quantitative analysis of Nrf2, TFRC, SLC7A11, GPX4, p21, and p16 levels, normalized to GAPDH (n = 4–7). (J) Intracellular ROS generation was measured using DCFH-DA, with ROS activity indicated by green fluorescence. Scale bar = 200 μm. (K) The quantitative analysis of ROS levels was conducted (n = 4). The data are presented as mean ± SD. Statistical significance is indicated as follows: *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001.

Techniques: CCK-8 Assay, Western Blot, Expressing, Activity Assay, Fluorescence

Journal: Frontiers in Pharmacology

Article Title: Targeted ErbB4 receptor activation prevents D-galactose-induced neuronal senescence via inhibiting ferroptosis pathway

doi: 10.3389/fphar.2025.1528604

Figure Lengend Snippet: ErbB4 receptor agonist mitigates D-gal-induced hippocampal neuronal aging both in vivo and in vitro through the activation of ErbB4 receptors and modulation of the Akt/Nrf2 signaling pathway. (A) Western blot analysis of pErbB4, ErbB4, pAkt1, and Akt1 protein expression levels in the hippocampus of mice. (B, C) Quantification of pErbB4 and pAkt1 levels (n = 4). (D) Western blot analysis of pErbB4, ErbB4, pAkt1, and Akt1 protein expression levels in D-gal-induced HT22 cells. (E, F) Quantification of pErbB4 and pAkt1 levels (n = 4). (G, H) Immunofluorescence analysis of the Nrf2-positive cells in the DG region (G) and in the cortex region (H) . Scale bar = 50 μm. (I) Western blot analysis of Nrf2, SLC7A11, GPX4, Lamin B1 and P21 protein levels after treatment of Nrf2 inhibitor AEM1. (J–N) Quantitative analysis of Nrf2, SLC7A11, GPX4, Lamin B1 and P21 levels (n = 4–7). Data are presented as mean ± SD. Statistical significance is indicated as follows: *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001.

Techniques: In Vivo, In Vitro, Activation Assay, Western Blot, Expressing, Immunofluorescence

Journal: Frontiers in Pharmacology

Article Title: Targeted ErbB4 receptor activation prevents D-galactose-induced neuronal senescence via inhibiting ferroptosis pathway

doi: 10.3389/fphar.2025.1528604

Figure Lengend Snippet: Schematic diagram illustrating the effects of targeted activation of ErbB4 receptor on neuronal aging via ferroptosis inhibition. Small molecule agonist (E4A)can activate ErbB4 receptors and regulate the Akt/Nrf2 signaling pathway to achieve neuroprotective effect in D-gal-induced neuronal aging. D-gal treatment increases the accumulation of advanced glycation end products (AGEs) which stimulates the production of reactive oxygen species (ROS) and increase the expression of senescence marker genes P53, P16, and P21. E4A can promote Akt1 phosphorylation and promote Nrf2 entrance into the nucleus, in which it involves in regulating the expression of ferroptosis suppressor gene, such as SLC7A11 and GPX4 to attenuate lipid peroxidation, which can inhibit cell ferroptosis and neuronal aging. The diagram was created with MedPeer ( www.medpeer.cn ).

Techniques: Activation Assay, Inhibition, Expressing, Marker

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

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: Other antibodies for the Western blot analysis, such as the rabbit anti-phospho-EGFR (Tyr1068), HER2 (2242s), phosphor-HER2 (Tyr1221/1222) (2243), phosphor-HER3 (Tyr1289) (4791), phosphor-HER4 (Tyr1284)/EGFR (Tyr1173) (4757), phosphor-MAPK (Tyr202/Tyr204) (4370), phospho-Akt (S473) (4060), phospho-STAT3 (Y705) (9145), phospho-SRC (Y416) (Tyr416) (6942) and β-actin (4970), were all obtained from Cell Signaling Technology Inc. (Hitchin, UK).

Techniques: Expressing, Cytometry, Fluorescence

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

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

Techniques: Colorimetric Assay, Polymer

Selectivity of receptor activation and downstream signaling by single ligands correlate in barcoded assays (A) Heatmap showing stimulation profiles on ERBB receptors, HTR2A, and downstream signaling pathways. Assays for receptors were performed using barcoded split TEV, assays for signaling pathways with pathway sensors coupled to barcodes. Compound effects are shown as log2-transformed fold change. (B–H) Barcoded assays align with luciferase readouts. Visualization of selected data from (A), comparing barcoded assays (black) with luciferase assay readouts (red). Assays for receptors were performed using split TEV, assays for signaling pathways with pathway sensors. Dose response graphs for EGFR (B), ERBB4 (B), EGR1p only (D), and EGR1p and ERBB4 transfected (E), HTR2A (F), CRE and HTR2A transfected (G), and NFAT and HTR2A transfected (H) with single stimuli applied at increasing concentrations. EGFld, EGF-like domain. Error bars represent SEM, n = 3 for barcoded assays, and n = 6 for luciferase assays. See also <xref ref-type=

Figure S2 and Table S2 . " width="100%" height="100%">

Journal: iScience

Article Title: Profiling of ERBB receptors and downstream pathways reveals selectivity and hidden properties of ERBB4 antagonists

doi: 10.1016/j.isci.2024.108839

Figure Lengend Snippet: Selectivity of receptor activation and downstream signaling by single ligands correlate in barcoded assays (A) Heatmap showing stimulation profiles on ERBB receptors, HTR2A, and downstream signaling pathways. Assays for receptors were performed using barcoded split TEV, assays for signaling pathways with pathway sensors coupled to barcodes. Compound effects are shown as log2-transformed fold change. (B–H) Barcoded assays align with luciferase readouts. Visualization of selected data from (A), comparing barcoded assays (black) with luciferase assay readouts (red). Assays for receptors were performed using split TEV, assays for signaling pathways with pathway sensors. Dose response graphs for EGFR (B), ERBB4 (B), EGR1p only (D), and EGR1p and ERBB4 transfected (E), HTR2A (F), CRE and HTR2A transfected (G), and NFAT and HTR2A transfected (H) with single stimuli applied at increasing concentrations. EGFld, EGF-like domain. Error bars represent SEM, n = 3 for barcoded assays, and n = 6 for luciferase assays. See also Figure S2 and Table S2 .

Article Snippet: Phosphorylation levels of EGFR and ERBB4 were assayed using p -EGFR-Y1068 (RRID: AB_2096270 ) (clone D7A5, dilution 1:500, No. 3777, Cell Signaling Technology) and p -ERBB4-Y1284 antibodies (RRID: AB_2099987 ) (clone 21A9, dilution 1:500, No. 4757, Cell Signaling Technology).

Techniques: Activation Assay, Protein-Protein interactions, Transformation Assay, Luciferase, Transfection

The barcoded ERBBprofiler reveals known and previously uncharacterized selectivity properties of ERBB receptor antagonists (A) Heatmap showing antagonistic effects of compounds on ERBB receptors, HTR2A, and downstream signaling pathways in PC12 cells. Assays for receptors were performed using barcoded split TEV assays, assays for signaling pathways with barcoded pathway sensors. In addition to the increasing concentrations of the compounds shown, all assays contained constant concentrations of EGF (30 ng/mL), EGF-like domain (10 ng/mL), and serotonin (1 μM). Compound effects are shown as log2-transformed fold change. (B–E) Dose response graphs comparing drug selectivity for receptors EGFR and ERBB4 (B, D) and downstream MAPK signaling (C, E) of compounds AG1478 (B, C), and pyrotinib (D, E). Data was extracted from the heatmap shown in (A). n = 3. (F–I) Dose response graphs for CRE sensor responses in PC12 cells using luciferase as readout for AG1478 (F), osimertinib (G), poziotinib (H), and pyrotinib (I). In addition to the increasing concentrations of the compounds shown, all assays contained the constant stimulation mix as in (A). (J–M) Dose response graphs for calcium and cAMP assays using Fluo-4 a.m. and GloSensor, respectively, as readouts in PC12 cells treated with increasing concentrations of AG1478 (J), osimertinib (K), poziotinib (L), and pyrotinib (M). As in luciferase assays, the constant stimulation mix was constantly present, next to the mentioned compounds. Error bars represent SEM, n = 3 for barcode assays (B-E), n = 6 for luciferase, Fluo-4 a.m., and GloSensor assays (F-M). See also <xref ref-type=

Figure S3 , Tables S3 and . " width="100%" height="100%">

Journal: iScience

Article Title: Profiling of ERBB receptors and downstream pathways reveals selectivity and hidden properties of ERBB4 antagonists

doi: 10.1016/j.isci.2024.108839

Figure Lengend Snippet: The barcoded ERBBprofiler reveals known and previously uncharacterized selectivity properties of ERBB receptor antagonists (A) Heatmap showing antagonistic effects of compounds on ERBB receptors, HTR2A, and downstream signaling pathways in PC12 cells. Assays for receptors were performed using barcoded split TEV assays, assays for signaling pathways with barcoded pathway sensors. In addition to the increasing concentrations of the compounds shown, all assays contained constant concentrations of EGF (30 ng/mL), EGF-like domain (10 ng/mL), and serotonin (1 μM). Compound effects are shown as log2-transformed fold change. (B–E) Dose response graphs comparing drug selectivity for receptors EGFR and ERBB4 (B, D) and downstream MAPK signaling (C, E) of compounds AG1478 (B, C), and pyrotinib (D, E). Data was extracted from the heatmap shown in (A). n = 3. (F–I) Dose response graphs for CRE sensor responses in PC12 cells using luciferase as readout for AG1478 (F), osimertinib (G), poziotinib (H), and pyrotinib (I). In addition to the increasing concentrations of the compounds shown, all assays contained the constant stimulation mix as in (A). (J–M) Dose response graphs for calcium and cAMP assays using Fluo-4 a.m. and GloSensor, respectively, as readouts in PC12 cells treated with increasing concentrations of AG1478 (J), osimertinib (K), poziotinib (L), and pyrotinib (M). As in luciferase assays, the constant stimulation mix was constantly present, next to the mentioned compounds. Error bars represent SEM, n = 3 for barcode assays (B-E), n = 6 for luciferase, Fluo-4 a.m., and GloSensor assays (F-M). See also Figure S3 , Tables S3 and .

Techniques: Protein-Protein interactions, Transformation Assay, Luciferase

Pyrotinib reveals selectivity for ERBB4 over EGFR (A and B) Dose response assays comparing AG1478 selectivity for receptors EGFR and ERBB4 (A) and downstream MAPK signaling (B) using firefly luciferase assays in PC12 cells. Assays for receptors were performed using split TEV, assays for MAPK signaling were conducted with an EGR1p pathway sensor. In addition to the increasing concentrations of AG1478, EGFR and ERBB4 assays contained a constant concentration of EGF (30 ng/mL) or EGF-like domain (10 ng/mL), respectively. (C and D) Western blot analyses of p -EGFR (in A549 cells) (C) and p -ERBB4 (in T-47 cells) (D) using increasing concentrations of AG1478. (E and F) Quantification of (C) and (D). (G and H) Dose response assays comparing pyrotinib selectivity for receptors EGFR and ERBB4 (G) and downstream MAPK signaling (H) using firefly luciferase assays in PC12 cells. Assays were conducted as in (A, B). (I and J) Western blot analyses of p -EGFR (in A549 cells) (I) and p -ERBB4 (in T-47 cells) (J) using increasing concentrations of pyrotinib. (K and L) Quantification of (I) and (J). Error bars represent SEM, n = 6 for luciferase assays (A, B, G, H), n = 3 for Western blot assays (E, F, K, L).

Journal: iScience

Article Title: Profiling of ERBB receptors and downstream pathways reveals selectivity and hidden properties of ERBB4 antagonists

doi: 10.1016/j.isci.2024.108839

Figure Lengend Snippet: Pyrotinib reveals selectivity for ERBB4 over EGFR (A and B) Dose response assays comparing AG1478 selectivity for receptors EGFR and ERBB4 (A) and downstream MAPK signaling (B) using firefly luciferase assays in PC12 cells. Assays for receptors were performed using split TEV, assays for MAPK signaling were conducted with an EGR1p pathway sensor. In addition to the increasing concentrations of AG1478, EGFR and ERBB4 assays contained a constant concentration of EGF (30 ng/mL) or EGF-like domain (10 ng/mL), respectively. (C and D) Western blot analyses of p -EGFR (in A549 cells) (C) and p -ERBB4 (in T-47 cells) (D) using increasing concentrations of AG1478. (E and F) Quantification of (C) and (D). (G and H) Dose response assays comparing pyrotinib selectivity for receptors EGFR and ERBB4 (G) and downstream MAPK signaling (H) using firefly luciferase assays in PC12 cells. Assays were conducted as in (A, B). (I and J) Western blot analyses of p -EGFR (in A549 cells) (I) and p -ERBB4 (in T-47 cells) (J) using increasing concentrations of pyrotinib. (K and L) Quantification of (I) and (J). Error bars represent SEM, n = 6 for luciferase assays (A, B, G, H), n = 3 for Western blot assays (E, F, K, L).

Techniques: Luciferase, Concentration Assay, Western Blot

The barcoded ERBBprofiler reveals novel ERBB4 selective antagonists (A) Heatmap showing antagonistic effects of LDC compounds on ERBB receptors, HTR2A, and downstream signaling pathways in PC12 cells. Assays for receptors were performed using barcoded split TEV assays, assays for signaling pathways using barcoded pathway sensors. In addition to the increasing concentrations of the compounds shown, all assays contained constant concentrations of EGF (30 ng/mL), EGF-like domain (10 ng/mL), and serotonin (1 μM). (B–E) Dose response graphs comparing drug selectivity for ERBB4 over EGFR (measured with split TEV) (B, C) and downstream MAPK signaling (measured with the EGR1p sensor) (D, E) of compound A (B, D), and compound B (C, E). Data was extracted from the heatmap shown in (A). (F–I) Orthogonal validation for compound B using Western blot analyses of p -EGFR and p -ERK1/2 (measured in A549 cells) (F) and p -ERBB4 and p -ERK1/2 (measured in T-47 cells) (G) using increasing concentrations of compound B (Cpd B). (H) Quantification of relative p -EGFR and p -ERBB4 from (F, G). (I) Quantification of relative p -ERK1/2 from (F, G). (J and K) In vitro kinase activity assays using LANCE assays for compound A (J) and compound B (K) showing dose response graphs comparing drug selectivity for ERBB4 (red) and EGFR (black). (L) Dose response graphs for CRE sensor responses in PC12 cells using luciferase as readout for compounds A (black) and B (red). In addition to the increasing concentrations of the compounds, the constant stimulation mix as in (A) was present. (M and N) Dose response graphs for calcium and cAMP assays using Fluo-4 a.m. (black) and GloSensor (red), respectively, as readouts in PC12 cells treated with increasing concentrations of compound A (M) and compound B (N). In addition to the compounds, the constant stimulation mix was present as in (A). Error bars represent SEM; n = 3 for barcode assays (B–E), Western blots (F–I) and in vitro kinase assays (J, K); n = 6 for luciferase assays, Fluo-4 a.m., and GloSensor assays (L–N). See also <xref ref-type=

Figures S4 and . " width="100%" height="100%">

Journal: iScience

Article Title: Profiling of ERBB receptors and downstream pathways reveals selectivity and hidden properties of ERBB4 antagonists

doi: 10.1016/j.isci.2024.108839

Figure Lengend Snippet: The barcoded ERBBprofiler reveals novel ERBB4 selective antagonists (A) Heatmap showing antagonistic effects of LDC compounds on ERBB receptors, HTR2A, and downstream signaling pathways in PC12 cells. Assays for receptors were performed using barcoded split TEV assays, assays for signaling pathways using barcoded pathway sensors. In addition to the increasing concentrations of the compounds shown, all assays contained constant concentrations of EGF (30 ng/mL), EGF-like domain (10 ng/mL), and serotonin (1 μM). (B–E) Dose response graphs comparing drug selectivity for ERBB4 over EGFR (measured with split TEV) (B, C) and downstream MAPK signaling (measured with the EGR1p sensor) (D, E) of compound A (B, D), and compound B (C, E). Data was extracted from the heatmap shown in (A). (F–I) Orthogonal validation for compound B using Western blot analyses of p -EGFR and p -ERK1/2 (measured in A549 cells) (F) and p -ERBB4 and p -ERK1/2 (measured in T-47 cells) (G) using increasing concentrations of compound B (Cpd B). (H) Quantification of relative p -EGFR and p -ERBB4 from (F, G). (I) Quantification of relative p -ERK1/2 from (F, G). (J and K) In vitro kinase activity assays using LANCE assays for compound A (J) and compound B (K) showing dose response graphs comparing drug selectivity for ERBB4 (red) and EGFR (black). (L) Dose response graphs for CRE sensor responses in PC12 cells using luciferase as readout for compounds A (black) and B (red). In addition to the increasing concentrations of the compounds, the constant stimulation mix as in (A) was present. (M and N) Dose response graphs for calcium and cAMP assays using Fluo-4 a.m. (black) and GloSensor (red), respectively, as readouts in PC12 cells treated with increasing concentrations of compound A (M) and compound B (N). In addition to the compounds, the constant stimulation mix was present as in (A). Error bars represent SEM; n = 3 for barcode assays (B–E), Western blots (F–I) and in vitro kinase assays (J, K); n = 6 for luciferase assays, Fluo-4 a.m., and GloSensor assays (L–N). See also Figures S4 and .

Techniques: Protein-Protein interactions, Biomarker Discovery, Western Blot, In Vitro, Activity Assay, Luciferase

Journal: iScience

Article Title: Profiling of ERBB receptors and downstream pathways reveals selectivity and hidden properties of ERBB4 antagonists

doi: 10.1016/j.isci.2024.108839

Figure Lengend Snippet:

Techniques: Virus, Recombinant, Biomarker Discovery, Software

Journal: iScience

Article Title: Profiling of ERBB receptors and downstream pathways reveals selectivity and hidden properties of ERBB4 antagonists

doi: 10.1016/j.isci.2024.108839

Figure Lengend Snippet:

Article Snippet: Rabbit monoclonal anti-phospho-HER4/ErbB4 (Tyr1284) (clone 21A9) , Cell Signaling Technology , Cat# 4757: RRID: AB_2099987.

Techniques: Virus, Recombinant, Biomarker Discovery, Software

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