Journal: bioRxiv

Article Title: Aberrant cortical development is driven by impaired cell cycle and translational control in a DDX3X syndrome model

doi: 10.1101/2022.02.21.481343

Figure Lengend Snippet: ( A ) RT-qPCR quantification of Ddx3x and Ddx3y mRNA levels in FACS-isolated Tdtomato+ cells from cKO male E11.5 cortices. ( B ) Schematic of Ddx3y CRISPR sgRNA electroporation of E14.5 brain. ( C ) RT-qPCR quantification of Ddx3y and Ddx3x levels in GFP+ FACS-isolated cells from E17.5 male and female mice electroporated with pCAG-GFP and either no sgRNA or Ddx3y sgRNA. ( D ) Representative sections of E17.5 male brains electroporated at E14.5 with pCAG-GFP and either no sgRNA or Ddx3y sgRNA and stained with anti-GFP (grey). Dotted lines, ventricular and pial surfaces; brackets delineate equivalently sized bins. ( E ) Quantification of distribution of GFP+ cells. ( F ) Same as ( D ), but sections were stained with anti-GFP (grey), Sox2 (magenta), and Neurod2 (red). ( G, H ) Quantification of GFP co-localization with Sox2 ( G ) or Neurod2 ( H ). Scale bars, indicated. Error bars, S.D. **p<0.01, ***p<0.001.

Article Snippet: The following primary antibodies were used, rat: anti-SOX2 (Thermo Fisher, 14-9811-82, 1:1000), anti-BrdU (Abcam, ab6326, 1:200); mouse: anti-TUJ1 (Biolegend, 801202, 1:2000), anti-CTIP2 (Abcam, c8035, 1:500); rabbit: anti-TBR2 (Abcam, AB23345, 1:1000), anti-CC3 (Cell Signaling, 9661, 1:250), anti-NEUROD2 (Abcam, AB104430, 1:500), anti-Ki67 (Cell Signaling Technology, 12202, 1:1000), anti-PH3 (Millipore, 06-570, 1:500), anti-TBR1 (Cell Signaling Technology, 49661S, 1:1000), anti-Lhx2 (Millipore, ABE1402, 1:500), anti-DDX3X (Sigma Aldrich, HPA001648, 1:500); chicken anti-GFP (Abcam, Ab13970, 1:1000).

Techniques: Quantitative RT-PCR, Isolation, CRISPR, Electroporation, Staining

Journal: bioRxiv

Article Title: Aberrant cortical development is driven by impaired cell cycle and translational control in a DDX3X syndrome model

doi: 10.1101/2022.02.21.481343

Figure Lengend Snippet: ( A ) RT- qPCR quantification of Setd3 knockdown in N2A cells. ( B ) Representative sections of E17.5 brains from mice electroporated at E14.5 with pCAG-GFP and scrambled or Setd3 siRNAs and immunostained with GFP (grey), Sox2 (magenta), Neurod2 (green). ( C ) Quantification of distribution of GFP-positive cells in 5 even bins of cortex. ( D, E ) Quantification of GFP co- localization with Sox2+ ( D ) and Neurod2+ ( E ) cells. ( F ) Schematic model summarizing how loss of DDX3X-dependent translation impairs neurogenesis. Scale bars, indicated. Error bars, S.D. *p<0.05, ****p<0.0001.

Article Snippet: The following primary antibodies were used, rat: anti-SOX2 (Thermo Fisher, 14-9811-82, 1:1000), anti-BrdU (Abcam, ab6326, 1:200); mouse: anti-TUJ1 (Biolegend, 801202, 1:2000), anti-CTIP2 (Abcam, c8035, 1:500); rabbit: anti-TBR2 (Abcam, AB23345, 1:1000), anti-CC3 (Cell Signaling, 9661, 1:250), anti-NEUROD2 (Abcam, AB104430, 1:500), anti-Ki67 (Cell Signaling Technology, 12202, 1:1000), anti-PH3 (Millipore, 06-570, 1:500), anti-TBR1 (Cell Signaling Technology, 49661S, 1:1000), anti-Lhx2 (Millipore, ABE1402, 1:500), anti-DDX3X (Sigma Aldrich, HPA001648, 1:500); chicken anti-GFP (Abcam, Ab13970, 1:1000).

Techniques: Quantitative RT-PCR

Journal: Behavioural Neurology

Article Title: Effects of Extremely Low-Frequency Electromagnetic Fields on Neurogenesis and Cognitive Behavior in an Experimental Model of Hippocampal Injury

doi: 10.1155/2017/9194261

Figure Lengend Snippet: Densitometric analysis of NeuroD2 bands (at approximately 48 kDa) in hippocampal extracts of the study groups. (a and b) Representative graph of NeuroD2 expression in different groups ( ∗ P < 0.05).

Article Snippet: The membranes were blocked with 5% skim milk in TBS, containing 0.5% Tween 20, and were incubated with primary antibodies directed against mouse monoclonal anti-NeuroD2 antibody (1 : 1500; Abcam, Cambridge, UK) and glyceraldehyde 3-phosphate dehydrogenase (GAPDH, 1 : 1000; Abcam, Cambridge, UK) for 2 h. After washing the membranes in a mixture of TBS and Tween 20 three times, they were reincubated with secondary alkaline-phosphatase-conjugated antibody (1 : 5000; Abcam, Cambridge, UK) for 1 h. Finally, the bands were detected with a chromogenic substrate (5-bromo-4-chloro-3-indolyl phosphate) in the presence of nitro blue tetrazolium.

Techniques: Expressing

Journal: BMC cardiovascular disorders

Article Title: Neurogenic differentiation 2 promotes inflammatory activation of macrophages in doxorubicin-induced myocarditis via regulating protein kinase D.

doi: 10.1186/s12872-025-04626-7

Figure Lengend Snippet: Fig. 4 Functional interaction between Neurogenic Differentiation 2 and protein kinase D. Macrophages were isolated from normal tissues and treated with 1µmol/L DOX for 12 h. (A) The binding site of PKD and Neurod2 was predicted by the JASPAR database. (B-C) qPCR analysis and western blotting were performed to detect the expression of Neurod2 in macrophages and tissues induced by DOX. (D-E) Macrophages isolated from the heart of normal mice were co-transfected with Neurod2 overexpression plasmid, and siNeurod2. qPCR and western blotting were performed to test the expression of Neurod2 mRNA and protein, respectively. (F-G) The levels of PKD in each group were explored by qPCR and western blotting. (H-I) The binding ability between PKD and Neurod2 was confirmed by dual-luciferase reporter assay (H) and CHIP assay (I). Values were presented as the mean ± SD of triplicate experiments. Compared with the control group or si-NC group, *P < 0.05, **P < 0. 01, ***P < 0. 001; Compared with the pc-NC group, #P < 0.05, ##P < 0. 01, ###P < 0. 001

Article Snippet: After blocking 1 h with 5% skimmed milk, the membranes were incubated with the following primary antibodies at 4°C overnight, including PKD (1:2000; Affinity Biosciences, Cincinnati, OH, USA), NLRP3 (1:2000; Affinity Biosciences), p-p65 (1:1000; Affinity Biosciences), p65 (1:1000; Affinity Biosciences), Neurod2 (1:1000; Biorbyt, Cambridge, UK.), GAPDH (1:3000; Affinity Biosciences).

Techniques: Functional Assay, Isolation, Binding Assay, Western Blot, Expressing, Transfection, Over Expression, Plasmid Preparation, Luciferase, Reporter Assay, Control

Journal: BMC cardiovascular disorders

Article Title: Neurogenic differentiation 2 promotes inflammatory activation of macrophages in doxorubicin-induced myocarditis via regulating protein kinase D.

doi: 10.1186/s12872-025-04626-7

Figure Lengend Snippet: Fig. 5 The overexpression of Protein kinase D can reverse the impact of neurogenic differentiation 2 on doxorubicin-induced inflammatory activation of macrophages. Macrophages isolated from normal tissue and treated with 1µmol/L DOX for 12 h. (A-B) Macrophages were co-transfected with PKD overexpression plasmid and si-Neurod2. qPCR was used to test the mRNA expressions of PKD and NLRP3. (C) The protein levels of PKD, p-p65, p65, and NLRP3 were detected. (D-G) The levels of TNF-α, IL-6, IL-18, and IL-1β in each group were measured by qPCR and ELISA. Values were presented as the mean ± SD of triplicate experiments. Compared with the DOX group, *P < 0.05, **P < 0. 01, ***P < 0. 001; Compared with DOX + pc-PKD group, #P < 0.05, ##P < 0. 01, ###P < 0. 001

Article Snippet: After blocking 1 h with 5% skimmed milk, the membranes were incubated with the following primary antibodies at 4°C overnight, including PKD (1:2000; Affinity Biosciences, Cincinnati, OH, USA), NLRP3 (1:2000; Affinity Biosciences), p-p65 (1:1000; Affinity Biosciences), p65 (1:1000; Affinity Biosciences), Neurod2 (1:1000; Biorbyt, Cambridge, UK.), GAPDH (1:3000; Affinity Biosciences).

Techniques: Over Expression, Activation Assay, Isolation, Transfection, Plasmid Preparation, Enzyme-linked Immunosorbent Assay

Journal: BMC cardiovascular disorders

Article Title: Neurogenic differentiation 2 promotes inflammatory activation of macrophages in doxorubicin-induced myocarditis via regulating protein kinase D.

doi: 10.1186/s12872-025-04626-7

Figure Lengend Snippet: Fig. 6 PKD-mediated macrophage inflammasome activation in heart tissue. The combination of NeuroD2 and PKD promoter could enhance NLRP3/ NF-κB signaling,

Article Snippet: After blocking 1 h with 5% skimmed milk, the membranes were incubated with the following primary antibodies at 4°C overnight, including PKD (1:2000; Affinity Biosciences, Cincinnati, OH, USA), NLRP3 (1:2000; Affinity Biosciences), p-p65 (1:1000; Affinity Biosciences), p65 (1:1000; Affinity Biosciences), Neurod2 (1:1000; Biorbyt, Cambridge, UK.), GAPDH (1:3000; Affinity Biosciences).

Techniques: Activation Assay

Journal: Immunity

Article Title: Prenatal interleukin 6 elevation increases glutamatergic synapse density and disrupts hippocampal connectivity in offspring

doi: 10.1016/j.immuni.2021.10.006

Figure Lengend Snippet:

Article Snippet: The following primary antibodies were used: guinea pig anti-VGLUT1 (1:1500; Synaptic Systems); mouse anti-V-gat (Synaptic Systems, 1:1000), rabbit anti-IBA (WAKO, 1:200), mouse anti-GFAP (Sigma-Aldrich, 1:400), guinea pig anti-NeuN (Synaptic System, 1:500), mouse anti-SATB2 (AbCam,1:100), rabbit anti-NeuroD2 (AbCam, 1:1500).

Techniques: Recombinant, Software