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: 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.

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: 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.

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: 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.

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: 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.

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: 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.

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: 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 ).

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: Activation Assay, Inhibition, Expressing, Marker