rabbit anti p erbb4  (Bioss)

Journal: Neurotherapeutics

Article Title: Targeted ErbB4 receptor activation ameliorates neuronal deficits via DOCK3 signaling in a transgenic mouse AD model

doi: 10.1016/j.neurot.2025.e00739

Figure Lengend Snippet: ErbB4 small molecule agonist ameliorates cognitive behavioral impairments in APP/PS1 mice. (A) Behavioral experimental scheme (D, days following the initiation of the behavioral experiment). (B) Representative traces in open field experiment. (C) The related statistical analysis of in open field experiment. (D) Representative traces in novel object recognition experiment. (E) The related statistical analyses in novel object recognition experiment. (F) Representative heat map in Y maze exploration experiment. (G) The rate of spontaneous alternation in Y maze experiment. (H) Representative swimming traces in water maze exploration experiment. (I) The one-way ANOVA statistical analysis of water maze exploration experiment. Tukey multiple comparative analyses were employed and data were presented as means ​± ​ ​standard error of the mean (SEM). ∗ p ​< ​0.05, ∗∗ p ​< ​0.01, and ∗∗∗ p ​< ​0.001. n ​= ​8 per group.

Article Snippet: Specifically, the fixed cells were treated with mouse anti-β3-tubulin antibody (1:100, sc80016, Santa Cruz biotech, USA) in combination with the following antibodies: rabbit anti-SYT1 (1:200, GB111130-100, Servicebio, Wuhan, China), rabbit anti-GAP43 antibody (1:1000, GB15094-100, Servicebio, Wuhan, China), rabbit anti- p -ErbB4 (1:200, bs-3220R, Bioss, Beijing, China), rabbit anti-SYP (1:200, AF8091, Bioss, Beijing, China), rabbit anti-PSD95 (1:200, GB11277-100, Servicebio, Wuhan, China) and rabbit anti-DOCK3 (1:200, 20683-1-AP, proteintech, Wuhan, China).

Techniques:

Journal: Neurotherapeutics

Article Title: Targeted ErbB4 receptor activation ameliorates neuronal deficits via DOCK3 signaling in a transgenic mouse AD model

doi: 10.1016/j.neurot.2025.e00739

Figure Lengend Snippet: ErbB4 small molecule agonist can improve the decrease of dendritic complexity and dendritic spine injury of hippocampal neurons in APP/PS1 mice. (A) Representative images of hippocampal neurons with a scale ​= ​50 ​μm; (B) dendrite length of hippocampal neurons; (C) The number of branching points of hippocampal neurons; (D) Representative images of neuronal dendritic segments, scale ​bar = ​10 ​μm; (E) total dendrite spine density of hippocampal neurons; (F) Mushroom spine density of hippocampal neurons. The results were presented as means ​± ​SEM. Significance levels were denoted as ∗ p ​< ​0.05, ∗∗ p ​< ​0.01, and ∗∗∗ p ​< ​0.001, with a sample size of n ​= ​4 per group.

Article Snippet: Specifically, the fixed cells were treated with mouse anti-β3-tubulin antibody (1:100, sc80016, Santa Cruz biotech, USA) in combination with the following antibodies: rabbit anti-SYT1 (1:200, GB111130-100, Servicebio, Wuhan, China), rabbit anti-GAP43 antibody (1:1000, GB15094-100, Servicebio, Wuhan, China), rabbit anti- p -ErbB4 (1:200, bs-3220R, Bioss, Beijing, China), rabbit anti-SYP (1:200, AF8091, Bioss, Beijing, China), rabbit anti-PSD95 (1:200, GB11277-100, Servicebio, Wuhan, China) and rabbit anti-DOCK3 (1:200, 20683-1-AP, proteintech, Wuhan, China).

Techniques:

Journal: Neurotherapeutics

Article Title: Targeted ErbB4 receptor activation ameliorates neuronal deficits via DOCK3 signaling in a transgenic mouse AD model

doi: 10.1016/j.neurot.2025.e00739

Figure Lengend Snippet: ErbB4 small molecule agonist can improve synaptic dysfunction of hippocampal neurons in APP/PS1 mice. (A) The ultrastructure of hippocampal neuron synapses was examined using transmission electron microscopy. (B) Statistical analysis was conducted on data related to hippocampal neuron synapses. (C) Levels of SYP and β3-tubulin detected by immunofluorescence. (D) Levels of DOCK3 and β3-tubulin detected by immunofluorescence. (E) Protein levels of the synapse-related proteins DOCK3, p -CREB, CREB, SYP, GAP43, PSD95, SYT1 in hippocampal neurons were assessed using Western blot analysis. (F) E statistical analysis was conducted on the data obtained from these protein levels, with Tukey multiple comparative analyses being utilized. Magnification, 10000 ​× ​for synaptic structure; scale bar, 1 ​μm. The results were presented as means ​± ​SEM. SYP: red; DOCK3: red; β3-tubulin: green; scale bar = 20 ​μm. Significance levels were denoted as ∗ p ​< ​0.05, ∗∗ p ​< ​0.01, and ∗∗∗ p ​< ​0.001, with a sample size of n ​= ​4–5 per group.

Article Snippet: Specifically, the fixed cells were treated with mouse anti-β3-tubulin antibody (1:100, sc80016, Santa Cruz biotech, USA) in combination with the following antibodies: rabbit anti-SYT1 (1:200, GB111130-100, Servicebio, Wuhan, China), rabbit anti-GAP43 antibody (1:1000, GB15094-100, Servicebio, Wuhan, China), rabbit anti- p -ErbB4 (1:200, bs-3220R, Bioss, Beijing, China), rabbit anti-SYP (1:200, AF8091, Bioss, Beijing, China), rabbit anti-PSD95 (1:200, GB11277-100, Servicebio, Wuhan, China) and rabbit anti-DOCK3 (1:200, 20683-1-AP, proteintech, Wuhan, China).

Techniques: Transmission Assay, Electron Microscopy, Immunofluorescence, Western Blot

Journal: Neurotherapeutics

Article Title: Targeted ErbB4 receptor activation ameliorates neuronal deficits via DOCK3 signaling in a transgenic mouse AD model

doi: 10.1016/j.neurot.2025.e00739

Figure Lengend Snippet: ErbB4 small molecule agonist can improve mitochondrial damage in hippocampal neurons of APP/PS1 mice. (A) The ultrastructure of hippocampal neuron mitochondria was examined using transmission electron microscopy. (B) Statistical analysis was conducted on data related to hippocampal neuron mitochondria. (C) Protein levels of the mitochondria-related proteins SIRT3 in hippocampal neurons were assessed using Western blot analysis. (D) Statistical analysis was conducted on the data obtained from these protein levels. (E) Levels of SIRT3 and β3-tubulin detected by immunofluorescence. SIRT3: red; β3-tubulin: green; scale bar = 20 ​μm. Tukey multiple comparative analyses were utilized. Magnification, 2000 ​× ​for mitochondrial structure; scale bar = 5 ​μm. The results were presented as means ​±SEM. Significance levels were denoted as ∗ p ​< ​0.05, ∗∗ p ​< ​0.01, and ∗∗∗ p ​< ​0.001, with a sample size of n ​= ​4–5 per group.

Article Snippet: Specifically, the fixed cells were treated with mouse anti-β3-tubulin antibody (1:100, sc80016, Santa Cruz biotech, USA) in combination with the following antibodies: rabbit anti-SYT1 (1:200, GB111130-100, Servicebio, Wuhan, China), rabbit anti-GAP43 antibody (1:1000, GB15094-100, Servicebio, Wuhan, China), rabbit anti- p -ErbB4 (1:200, bs-3220R, Bioss, Beijing, China), rabbit anti-SYP (1:200, AF8091, Bioss, Beijing, China), rabbit anti-PSD95 (1:200, GB11277-100, Servicebio, Wuhan, China) and rabbit anti-DOCK3 (1:200, 20683-1-AP, proteintech, Wuhan, China).

Techniques: Transmission Assay, Electron Microscopy, Western Blot, Immunofluorescence

Journal: Neurotherapeutics

Article Title: Targeted ErbB4 receptor activation ameliorates neuronal deficits via DOCK3 signaling in a transgenic mouse AD model

doi: 10.1016/j.neurot.2025.e00739

Figure Lengend Snippet: ErbB4 small molecule agonist can improve the ErbB4 signaling pathway in hippocampus of APP/PS1 mice. (A) Levels of p -ErbB4 and β3-tubulin detected by immunofluorescence. (B) Protein levels of the ErbB4 pathway proteins in hippocampal neurons were assessed using Western blot analysis. (C) Statistical analysis was conducted on the data obtained from these protein levels, with Tukey multiple comparative analyses being utilized. p -ErbB4: red; β3-tubulin: green; scale bar = 20 ​μm. The results were presented as means ​± ​SEM, with significance levels denoted as ∗ p ​< ​0.05, ∗∗ p ​< ​0.01, and ∗∗∗ p ​< ​0.001. Each experimental group consisted of n ​= ​5 samples.

Article Snippet: Specifically, the fixed cells were treated with mouse anti-β3-tubulin antibody (1:100, sc80016, Santa Cruz biotech, USA) in combination with the following antibodies: rabbit anti-SYT1 (1:200, GB111130-100, Servicebio, Wuhan, China), rabbit anti-GAP43 antibody (1:1000, GB15094-100, Servicebio, Wuhan, China), rabbit anti- p -ErbB4 (1:200, bs-3220R, Bioss, Beijing, China), rabbit anti-SYP (1:200, AF8091, Bioss, Beijing, China), rabbit anti-PSD95 (1:200, GB11277-100, Servicebio, Wuhan, China) and rabbit anti-DOCK3 (1:200, 20683-1-AP, proteintech, Wuhan, China).

Techniques: Immunofluorescence, Western Blot

Journal: Neurotherapeutics

Article Title: Targeted ErbB4 receptor activation ameliorates neuronal deficits via DOCK3 signaling in a transgenic mouse AD model

doi: 10.1016/j.neurot.2025.e00739

Figure Lengend Snippet: ErbB4 small molecule agonist can improve hippocampal neuroinflammation in APP/PS1 mice through TLR4/NLRP3 pathway. (A) Levels of Aβ and IBA1 detected by immunofluorescence staining. (B) Levels of Aβ and GFAP detected by immunofluorescence staining. (C) Western blot analysis was utilized to detect protein levels associated with the TLR4/NLRP3 pathway in microglial cells. (D) Statistical charts were generated to display the protein levels of TLR4/NLRP3. IBA1: red; GFAP: red; Aβ: green; scale bar = 50 ​μm. Tukey multiple comparative analyses were conducted, with data reported as means ​± ​SEM. Significance levels were denoted as ∗ p ​< ​0.05, ∗∗ p ​< ​0.01, ∗∗∗ p ​< ​0.001. Each group consisted of n ​= ​5 samples.

Article Snippet: Specifically, the fixed cells were treated with mouse anti-β3-tubulin antibody (1:100, sc80016, Santa Cruz biotech, USA) in combination with the following antibodies: rabbit anti-SYT1 (1:200, GB111130-100, Servicebio, Wuhan, China), rabbit anti-GAP43 antibody (1:1000, GB15094-100, Servicebio, Wuhan, China), rabbit anti- p -ErbB4 (1:200, bs-3220R, Bioss, Beijing, China), rabbit anti-SYP (1:200, AF8091, Bioss, Beijing, China), rabbit anti-PSD95 (1:200, GB11277-100, Servicebio, Wuhan, China) and rabbit anti-DOCK3 (1:200, 20683-1-AP, proteintech, Wuhan, China).

Techniques: Immunofluorescence, Staining, Western Blot, Generated

Journal: Neurotherapeutics

Article Title: Targeted ErbB4 receptor activation ameliorates neuronal deficits via DOCK3 signaling in a transgenic mouse AD model

doi: 10.1016/j.neurot.2025.e00739

Figure Lengend Snippet: ErbB4 small molecule agonist improves hippocampal neuronal injury in AD pathologic state in vitro through ErbB4 pathway. (A–F) After treatment with Aβ1-42 at 10 ​μM for 12 ​h, hippocampal neurons were treated with C11H7BrO3 at a concentration of 10 ​nM for an additional 24 ​h. (A) Protein levels of the ErbB4 pathway-related proteins in hippocampal neurons were assessed using Western blot analysis. (B) A statistical representation of these protein levels was generated in a chart. (C) Protein levels of SIRT3, CREB and GAP43 in hippocampal neurons were assessed using Western blot analysis. (D) Statistical representation of these protein levels was generated in a chart. (E) Protein levels of PSD95, SYP, SYT1, and DOCK3 in hippocampal neurons were assessed using Western blot analysis. (F) Statistical representation of these protein levels was generated in a chart. Tukey multiple comparative analyses were conducted, with data reported as means ​± ​SEM. Significance levels were denoted as ∗ p ​< ​0.05, ∗∗ p ​< ​0.01, ∗∗∗ p ​< ​0.001. A minimum sample size of n ​= ​5–6 per group was utilized.

Article Snippet: Specifically, the fixed cells were treated with mouse anti-β3-tubulin antibody (1:100, sc80016, Santa Cruz biotech, USA) in combination with the following antibodies: rabbit anti-SYT1 (1:200, GB111130-100, Servicebio, Wuhan, China), rabbit anti-GAP43 antibody (1:1000, GB15094-100, Servicebio, Wuhan, China), rabbit anti- p -ErbB4 (1:200, bs-3220R, Bioss, Beijing, China), rabbit anti-SYP (1:200, AF8091, Bioss, Beijing, China), rabbit anti-PSD95 (1:200, GB11277-100, Servicebio, Wuhan, China) and rabbit anti-DOCK3 (1:200, 20683-1-AP, proteintech, Wuhan, China).

Techniques: In Vitro, Concentration Assay, Western Blot, Generated

Journal: Neurotherapeutics

Article Title: Targeted ErbB4 receptor activation ameliorates neuronal deficits via DOCK3 signaling in a transgenic mouse AD model

doi: 10.1016/j.neurot.2025.e00739

Figure Lengend Snippet: The ErbB4 small molecule agonist modulates DOCK3 to ameliorate hippocampal neuronal injury under AD pathological conditions in vitro. (A–H) After a 12 ​h exposure to 10 ​μM ​Aβ, the cells were transfected with either 50 ​nM si-NC or si-DOCK3 for 24 ​h. This was followed by a combined treatment with C11H7BrO3 at a concentration of 10 ​nM for an additional 24 ​h. (A) Protein levels of the ErbB4 pathway-related proteins in hippocampal neurons were assessed using Western blot analysis. (B) Statistical representation of these protein levels was generated in a chart. (C) Protein levels of DOCK3, CREB, SIRT3 in hippocampal neurons were assessed using Western blot analysis. (D) Statistical representation of these protein levels was generated in a chart. (E) Protein levels of the synapse proteins SYT1, GAP43, SYP, and PSD95 in hippocampal neurons were assessed using Western blot analysis. (F) Statistical representation of these protein levels was generated in a chart. (G) Levels of oxidative stress indicators in hippocampal neurons, including T-SOD, GSH, GSH-Px, and MDA. (H) Intracellular ATP and extracellular ATP (eATP) levels in hippocampal neurons. (I–J). Representative images and quantitative analysis of mitochondrial signaling and membrane potential in hippocampal neurons. (K) The mitochondrial homeostasis function of hippocampal neurons involved the measurement of Mito-Tracker Green and TMRE levels. Tukey multiple comparative analyses were conducted, with data reported as means ​± ​SEM. Significance levels were denoted as ∗ p ​< ​0.05, ∗∗ p ​< ​0.01, ∗∗∗ p ​< ​0.001. Scale bar = 20 ​μm. A minimum sample size of n ​= ​5–6 per group was utilized.

Article Snippet: Specifically, the fixed cells were treated with mouse anti-β3-tubulin antibody (1:100, sc80016, Santa Cruz biotech, USA) in combination with the following antibodies: rabbit anti-SYT1 (1:200, GB111130-100, Servicebio, Wuhan, China), rabbit anti-GAP43 antibody (1:1000, GB15094-100, Servicebio, Wuhan, China), rabbit anti- p -ErbB4 (1:200, bs-3220R, Bioss, Beijing, China), rabbit anti-SYP (1:200, AF8091, Bioss, Beijing, China), rabbit anti-PSD95 (1:200, GB11277-100, Servicebio, Wuhan, China) and rabbit anti-DOCK3 (1:200, 20683-1-AP, proteintech, Wuhan, China).

Techniques: In Vitro, Transfection, Concentration Assay, Western Blot, Generated, Membrane

Journal: Neurotherapeutics

Article Title: Targeted ErbB4 receptor activation ameliorates neuronal deficits via DOCK3 signaling in a transgenic mouse AD model

doi: 10.1016/j.neurot.2025.e00739

Figure Lengend Snippet: ErbB4 small molecule agonist ameliorates cognitive behavioral impairments in APP/PS1 mice. (A) Behavioral experimental scheme (D, days following the initiation of the behavioral experiment). (B) Representative traces in open field experiment. (C) The related statistical analysis of in open field experiment. (D) Representative traces in novel object recognition experiment. (E) The related statistical analyses in novel object recognition experiment. (F) Representative heat map in Y maze exploration experiment. (G) The rate of spontaneous alternation in Y maze experiment. (H) Representative swimming traces in water maze exploration experiment. (I) The one-way ANOVA statistical analysis of water maze exploration experiment. Tukey multiple comparative analyses were employed and data were presented as means ​± ​ ​standard error of the mean (SEM). ∗ p ​< ​0.05, ∗∗ p ​< ​0.01, and ∗∗∗ p ​< ​0.001. n ​= ​8 per group.

Article Snippet: Specifically, the sections were treated with a mouse anti-β3-tubulin antibody (1:100, sc80016, Santa Cruz Biotechnology, USA) in combination with the following antibodies: rabbit anti- p -ErbB4 antibody (1:200, bs-3220R, Bioss, Beijing, China), rabbit anti-SYP antibody (1:200, AF8091, Bioss, Beijing, China), rabbit anti-SIRT3 antibody (1:200, bs6105R, Bioss, Beijing, China), and rabbit anti-DOCK3 antibody (1:200, 20683-1-AP, Proteintech, Wuhan, China).

Techniques:

Journal: Neurotherapeutics

Article Title: Targeted ErbB4 receptor activation ameliorates neuronal deficits via DOCK3 signaling in a transgenic mouse AD model

doi: 10.1016/j.neurot.2025.e00739

Figure Lengend Snippet: ErbB4 small molecule agonist can improve the decrease of dendritic complexity and dendritic spine injury of hippocampal neurons in APP/PS1 mice. (A) Representative images of hippocampal neurons with a scale ​= ​50 ​μm; (B) dendrite length of hippocampal neurons; (C) The number of branching points of hippocampal neurons; (D) Representative images of neuronal dendritic segments, scale ​bar = ​10 ​μm; (E) total dendrite spine density of hippocampal neurons; (F) Mushroom spine density of hippocampal neurons. The results were presented as means ​± ​SEM. Significance levels were denoted as ∗ p ​< ​0.05, ∗∗ p ​< ​0.01, and ∗∗∗ p ​< ​0.001, with a sample size of n ​= ​4 per group.

Article Snippet: Specifically, the sections were treated with a mouse anti-β3-tubulin antibody (1:100, sc80016, Santa Cruz Biotechnology, USA) in combination with the following antibodies: rabbit anti- p -ErbB4 antibody (1:200, bs-3220R, Bioss, Beijing, China), rabbit anti-SYP antibody (1:200, AF8091, Bioss, Beijing, China), rabbit anti-SIRT3 antibody (1:200, bs6105R, Bioss, Beijing, China), and rabbit anti-DOCK3 antibody (1:200, 20683-1-AP, Proteintech, Wuhan, China).

Techniques:

Journal: Neurotherapeutics

Article Title: Targeted ErbB4 receptor activation ameliorates neuronal deficits via DOCK3 signaling in a transgenic mouse AD model

doi: 10.1016/j.neurot.2025.e00739

Figure Lengend Snippet: ErbB4 small molecule agonist can improve synaptic dysfunction of hippocampal neurons in APP/PS1 mice. (A) The ultrastructure of hippocampal neuron synapses was examined using transmission electron microscopy. (B) Statistical analysis was conducted on data related to hippocampal neuron synapses. (C) Levels of SYP and β3-tubulin detected by immunofluorescence. (D) Levels of DOCK3 and β3-tubulin detected by immunofluorescence. (E) Protein levels of the synapse-related proteins DOCK3, p -CREB, CREB, SYP, GAP43, PSD95, SYT1 in hippocampal neurons were assessed using Western blot analysis. (F) E statistical analysis was conducted on the data obtained from these protein levels, with Tukey multiple comparative analyses being utilized. Magnification, 10000 ​× ​for synaptic structure; scale bar, 1 ​μm. The results were presented as means ​± ​SEM. SYP: red; DOCK3: red; β3-tubulin: green; scale bar = 20 ​μm. Significance levels were denoted as ∗ p ​< ​0.05, ∗∗ p ​< ​0.01, and ∗∗∗ p ​< ​0.001, with a sample size of n ​= ​4–5 per group.

Article Snippet: Specifically, the sections were treated with a mouse anti-β3-tubulin antibody (1:100, sc80016, Santa Cruz Biotechnology, USA) in combination with the following antibodies: rabbit anti- p -ErbB4 antibody (1:200, bs-3220R, Bioss, Beijing, China), rabbit anti-SYP antibody (1:200, AF8091, Bioss, Beijing, China), rabbit anti-SIRT3 antibody (1:200, bs6105R, Bioss, Beijing, China), and rabbit anti-DOCK3 antibody (1:200, 20683-1-AP, Proteintech, Wuhan, China).

Techniques: Transmission Assay, Electron Microscopy, Immunofluorescence, Western Blot

Journal: Neurotherapeutics

Article Title: Targeted ErbB4 receptor activation ameliorates neuronal deficits via DOCK3 signaling in a transgenic mouse AD model

doi: 10.1016/j.neurot.2025.e00739

Figure Lengend Snippet: ErbB4 small molecule agonist can improve mitochondrial damage in hippocampal neurons of APP/PS1 mice. (A) The ultrastructure of hippocampal neuron mitochondria was examined using transmission electron microscopy. (B) Statistical analysis was conducted on data related to hippocampal neuron mitochondria. (C) Protein levels of the mitochondria-related proteins SIRT3 in hippocampal neurons were assessed using Western blot analysis. (D) Statistical analysis was conducted on the data obtained from these protein levels. (E) Levels of SIRT3 and β3-tubulin detected by immunofluorescence. SIRT3: red; β3-tubulin: green; scale bar = 20 ​μm. Tukey multiple comparative analyses were utilized. Magnification, 2000 ​× ​for mitochondrial structure; scale bar = 5 ​μm. The results were presented as means ​±SEM. Significance levels were denoted as ∗ p ​< ​0.05, ∗∗ p ​< ​0.01, and ∗∗∗ p ​< ​0.001, with a sample size of n ​= ​4–5 per group.

Article Snippet: Specifically, the sections were treated with a mouse anti-β3-tubulin antibody (1:100, sc80016, Santa Cruz Biotechnology, USA) in combination with the following antibodies: rabbit anti- p -ErbB4 antibody (1:200, bs-3220R, Bioss, Beijing, China), rabbit anti-SYP antibody (1:200, AF8091, Bioss, Beijing, China), rabbit anti-SIRT3 antibody (1:200, bs6105R, Bioss, Beijing, China), and rabbit anti-DOCK3 antibody (1:200, 20683-1-AP, Proteintech, Wuhan, China).

Techniques: Transmission Assay, Electron Microscopy, Western Blot, Immunofluorescence

Journal: Neurotherapeutics

Article Title: Targeted ErbB4 receptor activation ameliorates neuronal deficits via DOCK3 signaling in a transgenic mouse AD model

doi: 10.1016/j.neurot.2025.e00739

Figure Lengend Snippet: ErbB4 small molecule agonist can improve the ErbB4 signaling pathway in hippocampus of APP/PS1 mice. (A) Levels of p -ErbB4 and β3-tubulin detected by immunofluorescence. (B) Protein levels of the ErbB4 pathway proteins in hippocampal neurons were assessed using Western blot analysis. (C) Statistical analysis was conducted on the data obtained from these protein levels, with Tukey multiple comparative analyses being utilized. p -ErbB4: red; β3-tubulin: green; scale bar = 20 ​μm. The results were presented as means ​± ​SEM, with significance levels denoted as ∗ p ​< ​0.05, ∗∗ p ​< ​0.01, and ∗∗∗ p ​< ​0.001. Each experimental group consisted of n ​= ​5 samples.

Article Snippet: Specifically, the sections were treated with a mouse anti-β3-tubulin antibody (1:100, sc80016, Santa Cruz Biotechnology, USA) in combination with the following antibodies: rabbit anti- p -ErbB4 antibody (1:200, bs-3220R, Bioss, Beijing, China), rabbit anti-SYP antibody (1:200, AF8091, Bioss, Beijing, China), rabbit anti-SIRT3 antibody (1:200, bs6105R, Bioss, Beijing, China), and rabbit anti-DOCK3 antibody (1:200, 20683-1-AP, Proteintech, Wuhan, China).

Techniques: Immunofluorescence, Western Blot

Journal: Neurotherapeutics

Article Title: Targeted ErbB4 receptor activation ameliorates neuronal deficits via DOCK3 signaling in a transgenic mouse AD model

doi: 10.1016/j.neurot.2025.e00739

Figure Lengend Snippet: ErbB4 small molecule agonist can improve hippocampal neuroinflammation in APP/PS1 mice through TLR4/NLRP3 pathway. (A) Levels of Aβ and IBA1 detected by immunofluorescence staining. (B) Levels of Aβ and GFAP detected by immunofluorescence staining. (C) Western blot analysis was utilized to detect protein levels associated with the TLR4/NLRP3 pathway in microglial cells. (D) Statistical charts were generated to display the protein levels of TLR4/NLRP3. IBA1: red; GFAP: red; Aβ: green; scale bar = 50 ​μm. Tukey multiple comparative analyses were conducted, with data reported as means ​± ​SEM. Significance levels were denoted as ∗ p ​< ​0.05, ∗∗ p ​< ​0.01, ∗∗∗ p ​< ​0.001. Each group consisted of n ​= ​5 samples.

Article Snippet: Specifically, the sections were treated with a mouse anti-β3-tubulin antibody (1:100, sc80016, Santa Cruz Biotechnology, USA) in combination with the following antibodies: rabbit anti- p -ErbB4 antibody (1:200, bs-3220R, Bioss, Beijing, China), rabbit anti-SYP antibody (1:200, AF8091, Bioss, Beijing, China), rabbit anti-SIRT3 antibody (1:200, bs6105R, Bioss, Beijing, China), and rabbit anti-DOCK3 antibody (1:200, 20683-1-AP, Proteintech, Wuhan, China).

Techniques: Immunofluorescence, Staining, Western Blot, Generated

Journal: Neurotherapeutics

Article Title: Targeted ErbB4 receptor activation ameliorates neuronal deficits via DOCK3 signaling in a transgenic mouse AD model

doi: 10.1016/j.neurot.2025.e00739

Figure Lengend Snippet: ErbB4 small molecule agonist improves hippocampal neuronal injury in AD pathologic state in vitro through ErbB4 pathway. (A–F) After treatment with Aβ1-42 at 10 ​μM for 12 ​h, hippocampal neurons were treated with C11H7BrO3 at a concentration of 10 ​nM for an additional 24 ​h. (A) Protein levels of the ErbB4 pathway-related proteins in hippocampal neurons were assessed using Western blot analysis. (B) A statistical representation of these protein levels was generated in a chart. (C) Protein levels of SIRT3, CREB and GAP43 in hippocampal neurons were assessed using Western blot analysis. (D) Statistical representation of these protein levels was generated in a chart. (E) Protein levels of PSD95, SYP, SYT1, and DOCK3 in hippocampal neurons were assessed using Western blot analysis. (F) Statistical representation of these protein levels was generated in a chart. Tukey multiple comparative analyses were conducted, with data reported as means ​± ​SEM. Significance levels were denoted as ∗ p ​< ​0.05, ∗∗ p ​< ​0.01, ∗∗∗ p ​< ​0.001. A minimum sample size of n ​= ​5–6 per group was utilized.

Article Snippet: Specifically, the sections were treated with a mouse anti-β3-tubulin antibody (1:100, sc80016, Santa Cruz Biotechnology, USA) in combination with the following antibodies: rabbit anti- p -ErbB4 antibody (1:200, bs-3220R, Bioss, Beijing, China), rabbit anti-SYP antibody (1:200, AF8091, Bioss, Beijing, China), rabbit anti-SIRT3 antibody (1:200, bs6105R, Bioss, Beijing, China), and rabbit anti-DOCK3 antibody (1:200, 20683-1-AP, Proteintech, Wuhan, China).

Techniques: In Vitro, Concentration Assay, Western Blot, Generated

Journal: Neurotherapeutics

Article Title: Targeted ErbB4 receptor activation ameliorates neuronal deficits via DOCK3 signaling in a transgenic mouse AD model

doi: 10.1016/j.neurot.2025.e00739

Figure Lengend Snippet: The ErbB4 small molecule agonist modulates DOCK3 to ameliorate hippocampal neuronal injury under AD pathological conditions in vitro. (A–H) After a 12 ​h exposure to 10 ​μM ​Aβ, the cells were transfected with either 50 ​nM si-NC or si-DOCK3 for 24 ​h. This was followed by a combined treatment with C11H7BrO3 at a concentration of 10 ​nM for an additional 24 ​h. (A) Protein levels of the ErbB4 pathway-related proteins in hippocampal neurons were assessed using Western blot analysis. (B) Statistical representation of these protein levels was generated in a chart. (C) Protein levels of DOCK3, CREB, SIRT3 in hippocampal neurons were assessed using Western blot analysis. (D) Statistical representation of these protein levels was generated in a chart. (E) Protein levels of the synapse proteins SYT1, GAP43, SYP, and PSD95 in hippocampal neurons were assessed using Western blot analysis. (F) Statistical representation of these protein levels was generated in a chart. (G) Levels of oxidative stress indicators in hippocampal neurons, including T-SOD, GSH, GSH-Px, and MDA. (H) Intracellular ATP and extracellular ATP (eATP) levels in hippocampal neurons. (I–J). Representative images and quantitative analysis of mitochondrial signaling and membrane potential in hippocampal neurons. (K) The mitochondrial homeostasis function of hippocampal neurons involved the measurement of Mito-Tracker Green and TMRE levels. Tukey multiple comparative analyses were conducted, with data reported as means ​± ​SEM. Significance levels were denoted as ∗ p ​< ​0.05, ∗∗ p ​< ​0.01, ∗∗∗ p ​< ​0.001. Scale bar = 20 ​μm. A minimum sample size of n ​= ​5–6 per group was utilized.

Article Snippet: Specifically, the sections were treated with a mouse anti-β3-tubulin antibody (1:100, sc80016, Santa Cruz Biotechnology, USA) in combination with the following antibodies: rabbit anti- p -ErbB4 antibody (1:200, bs-3220R, Bioss, Beijing, China), rabbit anti-SYP antibody (1:200, AF8091, Bioss, Beijing, China), rabbit anti-SIRT3 antibody (1:200, bs6105R, Bioss, Beijing, China), and rabbit anti-DOCK3 antibody (1:200, 20683-1-AP, Proteintech, Wuhan, China).

Techniques: In Vitro, Transfection, Concentration Assay, Western Blot, Generated, Membrane

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

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 .

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: Protein-Protein interactions, Transformation 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: 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).

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: Luciferase, Concentration Assay, Western Blot

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 .

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: 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:

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: Virus, Recombinant, Biomarker Discovery, Software