Journal: The Journal of Neuroscience

Article Title: Neuregulin1 Nuclear Signaling Influences Adult Neurogenesis and Regulates a Schizophrenia Susceptibility Gene Network within the Mouse Dentate Gyrus

doi: 10.1523/JNEUROSCI.0063-24.2024

Figure Lengend Snippet: Key resources table

Article Snippet: Recombinant mouse ErbB4/Her4 Fc chimera protein, CF , R&D Systems , Catalog #4387-ER-050.

Techniques: Affinity Purification, Virus, Recombinant, Protease Inhibitor, Sequencing, RNAscope, Variant Assay, Plasmid Preparation, Software

Journal: The Journal of Neuroscience

Article Title: Neuregulin1 Nuclear Signaling Influences Adult Neurogenesis and Regulates a Schizophrenia Susceptibility Gene Network within the Mouse Dentate Gyrus

doi: 10.1523/JNEUROSCI.0063-24.2024

Figure Lengend Snippet: The V 321 L substitution decreases nuclear back signaling. A , Immunoblot of triplicate nuclear fractions isolated from pooled cortical and hippocampal lysates. Nrg1 ICD was detected using Santa Cruz Biotechnology antibody sc-348. Histone H3 served as a nuclear loading control and Na+/K+ ATPase as a marker for the membrane fraction ( N = 3 mice/genotype). B , Immunoblot of two replicates of membrane fractions isolated from pooled cortical and hippocampal lysates. NRG1 ICD was detected using Santa Cruz Biotechnology antibody sc-348. Na+/K+ ATPase served as a marker for the membrane fraction; note the lack of the nuclear marker Histone H3 or cytoplasmic marker CyclophilinA indicating clean membrane preps. FL Nrg1 is indicated with a yellow arrowhead as “Nrg1-FL,” and the membrane-bound C-terminal fragment not cleaved by γ-secretase is indicated as “Nrg1 TM-CTF.” A positive control consisting of total lysate from N2A cells transfected with a Type III Nrg1 plasmid is shown in the lane labeled “C.” ( N = 2 mice/genotype.) C , Left, Hippocampal neurons from WT (dark blue) and V 321 L (light blue) neonatal pups (P4) were cultured for 17 d in vitro and were stimulated with either vehicle (Veh), 20 nM sERBB4 (sB4), or 20 nM sErbB4 after a 24 h pretreatment with 20 µM of the γ-secretase inhibitor DAPT (DAPT). Neurons were fixed and stained using an antibody directed against the Nrg1 ICD and counterstained with DAPI. Scale bar, 10 µm. Right, Quantification of nuclear clusters of Nrg1-ICD. Neurons from WT mice show increased nuclear ICD clusters in response to sB4 stimulation, which is counteracted by pretreatment with DAPT (DAPT). Neurons from V 321 L mice do not respond to sB4 stimulation ( N = 6–13 neurons, 3 platings/mouse, 3 mice/genotype; one-way ANOVA p values corrected for multiple comparisons using Tukey’s post hoc test; WT Veh vs WT sB4, p < 0.0001 (****); WT sB4 vs WT sB4 + DAPT, **** p < 0.0001; WT sB4 vs V 321 L Veh, **** p < 0.0001; WT sB4 vs V 321 L B4, **** p < 0.0001). All other comparisons are statistically not statistically significant. D , Cortical neurons from embryonic WT (dark blue) and V 321 L mice (light blue; E18.5) were cultured for DIV3 and were stimulated with soluble ErbB4 (sB4), PI3K inhibitor WM, γ-secretase inhibitor L-685,458 (L6), WM + B4, or L6 + B4. Neurons that underwent no drug treatments/sB4 stimulation are indicated as the control group (C). Neurons were fixed and axonal length was quantified. (Two-way ANOVA with Tukey’s post hoc correction, WT C vs WT B4, **** p = 0.0002; WT L6 vs WT L6 + B4, ** p = 0.0047; V 321 L C vs V 321 L B4, *** p = 0.001; V 321 L WM vs V 321 L WM + B4, p = 0.1; V 321 L L6 vs V 321 L L6 + B4, * p = 0.03.) N = 20–37 neurons per genotype per condition. ns, not significant. E , Treatment and conditions same as in D . Quantification is for dendritic length. (two-way ANOVA w/ Tukey’s post hoc correction: WT C vs WT B4, ** p = 0.002; WT WM vs WT WM + B4, **** p < 0.0001). N = 20–37 neurons per genotype per condition. ns, not significant.

Article Snippet: Recombinant mouse ErbB4/Her4 Fc chimera protein, CF , R&D Systems , Catalog #4387-ER-050.

Techniques: Western Blot, Isolation, Control, Marker, Membrane, Positive Control, Transfection, Plasmid Preparation, Labeling, Cell Culture, In Vitro, Staining

Journal: Cancer control : journal of the Moffitt Cancer Center

Article Title: ERBB4 Expression in Ovarian Serous Carcinoma Resistant to Platinum-Based Therapy

doi: 10.1177/107327481702400115

Figure Lengend Snippet: Mean level of ERBB4 expression in normal controls, tumor tissue, and ovarian cancer cell lines. Analysis of variance was statistically significant (P < .001). The mean scores of ERBB4 are significantly different for tumors vs controls vs cell lines, with cell lines greater than tumors and greater than controls.

Article Snippet: A mouse monoclonal antibody that reacts to ERBB4, NB100–2662 (Novus, Littleton, CO), was used at a 1:25 concentration in Antibody Diluent (Dako, Carpenteria, CA) and incubated for 60 minutes.

Techniques: Expressing

Journal: Cancer control : journal of the Moffitt Cancer Center

Article Title: ERBB4 Expression in Ovarian Serous Carcinoma Resistant to Platinum-Based Therapy

doi: 10.1177/107327481702400115

Figure Lengend Snippet: ERBB4 expression by immunohistochemistry. Shown are (A) rare membranous, (B) nuclear staining, (C–G) increasing H scores from less than 50 to 300 for particular fields, and (H) ERBB4 staining in cells lines.

Article Snippet: A mouse monoclonal antibody that reacts to ERBB4, NB100–2662 (Novus, Littleton, CO), was used at a 1:25 concentration in Antibody Diluent (Dako, Carpenteria, CA) and incubated for 60 minutes.

Techniques: Expressing, Immunohistochemistry, Staining

Journal: Cancer control : journal of the Moffitt Cancer Center

Article Title: ERBB4 Expression in Ovarian Serous Carcinoma Resistant to Platinum-Based Therapy

doi: 10.1177/107327481702400115

Figure Lengend Snippet: Using an unpaired t test with Welch correction, ERBB4 expression was higher patients with an IR than CR to platinum-based therapy (P = .033).

Article Snippet: A mouse monoclonal antibody that reacts to ERBB4, NB100–2662 (Novus, Littleton, CO), was used at a 1:25 concentration in Antibody Diluent (Dako, Carpenteria, CA) and incubated for 60 minutes.

Techniques: Expressing

Journal: Cancer control : journal of the Moffitt Cancer Center

Article Title: ERBB4 Expression in Ovarian Serous Carcinoma Resistant to Platinum-Based Therapy

doi: 10.1177/107327481702400115

Figure Lengend Snippet: Multivariate Survival Analysis Using a Cox Proportional Hazard Model

Article Snippet: A mouse monoclonal antibody that reacts to ERBB4, NB100–2662 (Novus, Littleton, CO), was used at a 1:25 concentration in Antibody Diluent (Dako, Carpenteria, CA) and incubated for 60 minutes.

Techniques:

Journal: Cancer control : journal of the Moffitt Cancer Center

Article Title: ERBB4 Expression in Ovarian Serous Carcinoma Resistant to Platinum-Based Therapy

doi: 10.1177/107327481702400115

Figure Lengend Snippet: Log-rank test results illustrating ERBB4 expression and OS. Probability values were as follows: all 3 levels, P = .020; low vs intermediate, P = .080; low vs high, P = .004; and intermediate vs high, P = .201.

Article Snippet: A mouse monoclonal antibody that reacts to ERBB4, NB100–2662 (Novus, Littleton, CO), was used at a 1:25 concentration in Antibody Diluent (Dako, Carpenteria, CA) and incubated for 60 minutes.

Techniques: Expressing