- MA5-33103 - Invitrogen Antibodies RBFOX3 antibody

Submitted references Icarisid II rescues cognitive dysfunction via activation of Wnt/β-catenin signaling pathway promoting hippocampal neurogenesis in APP/PS1 transgenic mice.

Xiao HH, Chen JC, Li H, Li RH, Wang HB, Song HP, Li HY, Shan GS, Tian Y, Zhao YM, Tian JM, Yang JX
Phytotherapy research : PTR 2022 May;36(5):2095-2108

Use of high-content imaging to quantify transduction of AAV-PHP viruses in the brain following systemic delivery.

Smith EJ, Farshim PP, Flomen R, Jones ST, McAteer SJ, Deverman BE, Gradinaru V, Bates GP
Brain communications 2021;3(2):fcab105

Active constituent of Polygala tenuifolia attenuates cognitive deficits by rescuing hippocampal neurogenesis in APP/PS1 transgenic mice.

Wang XF, Xiao HH, Wu YT, Kong L, Chen JC, Yang JX, Hu XL
BMC complementary medicine and therapies 2021 Oct 25;21(1):267

UNC-45A Is Highly Expressed in the Proliferative Cells of the Mouse Genital Tract and in the Microtubule-Rich Areas of the Mouse Nervous System.

Clemente V, Hoshino A, Meints J, Shetty M, Starr T, Lee M, Bazzaro M
Cells 2021 Jun 26;10(7)

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Supportive validation

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Experimental details: Western blot of NeuN in rat cortical lysate showing specific immunolabeling of a ~46/48 kDa band corresponding to NeuN monoclonal antibody (Product # MA5-33103).

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Experimental details: Immunocytochemistry analysis of NeuN in cultured rat neurons showing strong nuclear and distal cytoplasmic labeling with NeuN monoclonal antibody (Product # MA5-33103) using a dilution of 1:1,000 (red). The complete absence of astrocyte staining is shown using Anti-GFAP with a dilution of 1:1,000 (green) and nuclear staining was done with DAPI (blue).

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Experimental details: Immunohistochemistry analysis of NeuN in rat brain stem co-labeled with NeuN monoclonal antibody (Product # MA5-33103) using a dilution of 1:1,000 (green) and Anti-MAP2 using a dilution of 1:5,000 (red). The Anti-FOX3 specifically labels the nuclei and the proximal cytoplasm of neuronal cells while the Anti-MAP2 labels dendrites and overlaps with FOX3 labeling the perikarya of neurons. The blue is DAPI staining of nuclear DNA.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Figure 2 Building an automated analysis tree to quantify AAV transduction . ( A ) Brain sections were stained with Hoechst and immunostained for NeuN and GFP and images were captured on the Opera Phenix. Whole brain composites were generated by stitching together a panel of images with a 10% overlap. This was performed for 19 optical slices captured throughout the Z-plane at 1.2 mum intervals. ( B ) Each channel was analysed separately to identify positive signals for cell nuclei (Hoechst, blue), neurons (NeuN, orange) and AAV transduced cells (GFP, green). The Harmony software pseudo-labels positive signals with a coloured border as shown in the lower right-hand side of the thumbnails. ( C ) Positive signals for NeuN and GFP were further gated based on user input and machine learning to remove false positives. A combination of these gated outputs were used to identify NeuN+, GFP+ and NeuN+/GFP+ cells. ( D ) Data from each optical slice throughout the Z-stack were combined to locate neurons and transduced cells throughout the entire section. ( E-G ) The percentage of the Hoechst+ cells found in each section that were NeuN+, GFP+ or both NeuN+/GFP+ for each mouse strain. (H) The automated analysis of the data collected from all brain sections was reanalyzed using less stringent gating criteria for GFP+. Allowing more cells to be registered as GFP+ increased the transduction efficiency in the C57BL/6J mice. However, a small percentage of the Hoechst+ cells in the CBA/Ca section

Supportive validation

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Experimental details: Fig. 9 DISS promoted the neuronal differentiation of hippocampal NSCs in APP/PS1 mice. IF analysis against NeuN was carried out to detect the mature NSCs in the hippocampus of APP/PS1 mice. (A, C and E) Representative immunostaining images of NeuN + cells in the hippocampal CA1 (A), CA3 (C) and DG (E) regions, respectively. Scale bar = 100 mum. (B, D and F). Quantitative analysis of NeuN + cells in the hippocampal CA1 (B), CA3 (D) and DG (F) regions, respectively. # P < 0.05; ## P < 0.01 vs. WT group; * P < 0.05 vs. APP/PS1 group. Data are represented as mean +- sem. n = 6 mice in each group; 6 slices per animal were analyzed by one-way ANOVA

Supportive validation

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Experimental details: 8 FIGURE ICS II promoted the neuronal differentiation of NSCs in the hippocampus by activation of Wnt/beta-actenin signaling pathway. Mice were administrated with ICS II (10 mg/kg) accompanied with or without XAV-939 (5 mg/kg) for 4 weeks, followed by intraperitoneal injection of BrdU (100 mg/kg) to label the proliferation cells. After treatment for another 3 weeks, immunostaining against NeuN and BrdU were carried out to detect the mature neurons derived from proliferative NSCs in the hippocampal DG regions. (a) Representative images of NeuN + /BrdU + cells; (b) Quantification of NeuN + cells as shown in A; (c) Quantification of NeuN + /BrdU + cells as shown in A. Data are represented as mean +- SEM ( n = 3/group). ## p < .01 versus WT group; ** p < .01 versus TG group; && p < .01 versus ICS II group; n.s p > .05 versus TG group. ANOVA followed by Tukey's post-hoc test was used

MA5-33103

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