Journal: International journal of molecular sciences

Article Title: Effectiveness of Combinational Treatments for Alzheimer's Disease with Human Neural Stem Cells and Microglial Cells Over-Expressing Functional Genes.

doi: 10.3390/ijms24119561

Figure Lengend Snippet: Figure 1. Expression of mRNAs and production of functional proteins in human neural stem cells and microglial cells, and amyloid-β (Aβ) clearance by microglial cells. (A) RT-PCR analysis of ChAT, NEP and SRA mRNAs. (B) Immunocytochemical staining on ChAT, NEP and SRA (green). (C) Clearance of Aβ peptides by HMO6 (#), HMO6.NEP (•) and HMO6.SRA (▼). * Significantly different from HMO6 cells at each time point (p < 0.05).

Article Snippet: Immunohistochemical Analysis of hMito and Functional Proteins The mouse brains were perfusion-fixed with 4% paraformaldehyde solution and postfixed for 48 h, followed by cryoprotection in 30% sucrose for 72 h. Coronal cryosections in 30-μm thickness were prepared and processed for double immunostaining of human mitochondria (hMito) and/or ChAT, NEP or SRA using antibodies specific for hMito (1:100, Millipore), ChAT (1:100, Invitrogen), NEP (1:100, Invitrogen) and SRA (1:100, Bioss).

Techniques: Expressing, Functional Assay, Reverse Transcription Polymerase Chain Reaction, Staining

Journal: International journal of molecular sciences

Article Title: Effectiveness of Combinational Treatments for Alzheimer's Disease with Human Neural Stem Cells and Microglial Cells Over-Expressing Functional Genes.

doi: 10.3390/ijms24119561

Figure Lengend Snippet: Figure 4. Recovery of learning and memory functions four weeks post-injection in passive avoid- ance (A) Morris water-maze (B) studies, and tracking of water-maze swimming performance (C). #: Normal, •: AF64A alone, ▼: F3, △: F3.ChAT, ■: HMO6, □: HMO6.NEP, ◆: HMO6.SRA, 3: F3.ChAT + HMO6.NEP, ▲: F3.ChAT + HMO6.SRA. * Significantly different from normal (p < 0.05). # Significantly different from AF64A alone (p < 0.05).

Article Snippet: Immunohistochemical Analysis of hMito and Functional Proteins The mouse brains were perfusion-fixed with 4% paraformaldehyde solution and postfixed for 48 h, followed by cryoprotection in 30% sucrose for 72 h. Coronal cryosections in 30-μm thickness were prepared and processed for double immunostaining of human mitochondria (hMito) and/or ChAT, NEP or SRA using antibodies specific for hMito (1:100, Millipore), ChAT (1:100, Invitrogen), NEP (1:100, Invitrogen) and SRA (1:100, Bioss).

Techniques: Injection

Journal: International journal of molecular sciences

Article Title: Effectiveness of Combinational Treatments for Alzheimer's Disease with Human Neural Stem Cells and Microglial Cells Over-Expressing Functional Genes.

doi: 10.3390/ijms24119561

Figure Lengend Snippet: Figure 5. Immunohistochemical identification of human neural stem cells (F3) and microglial cells (HMO6) and functional proteins (ChAT, NEP and SRA) via double immunostaining for human mitochondria (hMito) and ChAT, NEP or SRA.

Article Snippet: Immunohistochemical Analysis of hMito and Functional Proteins The mouse brains were perfusion-fixed with 4% paraformaldehyde solution and postfixed for 48 h, followed by cryoprotection in 30% sucrose for 72 h. Coronal cryosections in 30-μm thickness were prepared and processed for double immunostaining of human mitochondria (hMito) and/or ChAT, NEP or SRA using antibodies specific for hMito (1:100, Millipore), ChAT (1:100, Invitrogen), NEP (1:100, Invitrogen) and SRA (1:100, Bioss).

Techniques: Immunohistochemical staining, Functional Assay, Double Immunostaining

Journal: Neurobiology of disease

Article Title: Altered SYNJ2BP-mediated mitochondrial-ER contacts in motor neuron disease

doi: 10.1016/j.nbd.2022.105832

Figure Lengend Snippet: SYNJ2BP levels are increased in spinal cord tissue of patients with motor neuron disease A Three SBMA (labeled SB1, SB2, and SB3), and two ALS4 (labeled ALS4–1 and ALS4–2) autopsy patient samples were used for this study. The age of onset of SBMA patients ranged from 39 to 67 years old with disease duration ranging from 16 to 30 years. The diagnosis of SBMA and ALS4 were established by clinical features and confirmatory genetic testing. FHx: Family history, EMG: Electromyography. B SYNJ2BP gene expression by RT-PCR using post-mortem spinal cord tissue. Results represent average fold change from three independent experiments, each experiment included N = 3 controls, N = 3 SBMA patients, and N = 2 ALS4 patients. C Representative western blot image showing increased SYNJ2BP levels in SBMA and ALS4 patient spinal cord, and a reduction in ChAT. Actin was used as a loading control. D Quantification of western blot analysis. SYNJ2BP signal intensity was normalized to the ChAT signal. P value not shown for comparison to ALS4 spinal cord as only two samples were analyzed. E IHC staining of SYNJ2BP in FFPE spinal cord tissue from an SBMA patient. Scale bar 100 μm. F SYNJ2BP signal intensity normalized to cell surface area using ImageJ. Data represent individual motor neurons. All motor neurons within each tissue section were counted. Unpaired t -test, error bars represent ± SEM; *p < 0.05, **p < 0.01, and *** p < 0.001.

Article Snippet: The Taqman assays used included ChAT (Hs00758143_m1), SYNJ2BP (Hs00218058_m1), and Actin (Hs00157387_m1).

Techniques: Labeling, Biomarker Discovery, Gene Expression, Reverse Transcription Polymerase Chain Reaction, Western Blot, Control, Comparison, Immunohistochemistry

Journal: Neurobiology of disease

Article Title: Altered SYNJ2BP-mediated mitochondrial-ER contacts in motor neuron disease

doi: 10.1016/j.nbd.2022.105832

Figure Lengend Snippet: Increased MERC in SBMA spinal motor neurons. A PLA combined with ChAT immunofluorescent staining shows increased VDAC1 interaction with IP3R in SBMA patients spinal cord tissue and, B SYNJ2BP interaction with its known interacting partner RRBP1. Arrowheads pointing at the PLA signal (red) in the motor neurons (green), scale bar 20 μm. Zoom of the boxed region shows increased PLA signal in a representative motor neuron.

Article Snippet: The Taqman assays used included ChAT (Hs00758143_m1), SYNJ2BP (Hs00218058_m1), and Actin (Hs00157387_m1).

Techniques: Staining