Journal: Life Medicine

Article Title: CRISPR/Cas9-mediated genetic correction reverses spinocerebellar ataxia 3 disease-associated phenotypes in differentiated cerebellar neurons

doi: 10.1093/lifemedi/lnac020

Figure Lengend Snippet: Reversion of SCA3 disease-associated phenotypes in genetically corrected cerebellar neurons derived from cerebellar NSCs.(A) The phase contrast of cerebellar neurons from cerebellar NSCs after prolonged differentiation (>6 weeks) upon growth factor-withdrawal. (B) Expression of MAP2 + neurons and GFAP + glia in 6-week-differentiated neural cultures. (C and D) Immunostaining for L7, CALB1 and TUJ1 on cultured cerebellar neurons (differentiation >6 weeks). (E–H) Immunostaining on 6-week-differentiated neural cultures for granule cells markers (ATOH1, PAX6 and ZIC2). (I and J) The large majority of cerebellar NSC-derived neurons display a glutamatergic phenotype, staining positively for VGLUT1 (J). Only occasional neurons exhibit GABAergic phenotypes (I). (K) Percentage of cells positive for TUJ1, MAP2, GFAP, CALB1, L7, ATOH1 and PAX6 in 6-week-differentiated SCA3, corrected and WT cerebellar neurons. Each bar represents mean ± SD with three biological replicates. (L–P) TUNEL staining and the percentage of apoptotic cells in SCA3, corrected and WT cerebellar neurons after prolonged differentiation (>6 weeks) with or without nutritional factors (NF), which were supplemented for growth and long-term viability of neurons. Each bar represents mean ± SD with three biological replicates. (Q) Cell proliferation was assayed in 3-week-differentiated cultures derived from SCA3, corrected and WT NSCs with or without NF using a BrdU cell proliferation assay kit. Each bar represents mean ± SD with three biological replicates. +NF, normal culture condition supplemented with nutritional factors; ‐NF, nutritional factor-withdrawal condition. Samples included SCA3 (Pa1 and Pa2), corrected (C-21 and 1A) and WT (WT1 and WT2) neurons. The scale bars represent 500 μm (A), 20 μm (B, D, F and I) and 50 μm (C, E, J, G, H and L–O). *** P < 0.001 was determined by unpaired t test. n.s., not significant.

Article Snippet: VGLUT1 , Proteintech , Rabbit , 50.

Techniques: Derivative Assay, Expressing, Immunostaining, Cell Culture, Staining, TUNEL Assay, BrdU Cell Proliferation Assay

Journal: Life Medicine

Article Title: CRISPR/Cas9-mediated genetic correction reverses spinocerebellar ataxia 3 disease-associated phenotypes in differentiated cerebellar neurons

doi: 10.1093/lifemedi/lnac020

Figure Lengend Snippet:

Article Snippet: VGLUT1 , Proteintech , Rabbit , 50.

Techniques:

Journal: eNeuro

Article Title: Synaptic Organization of VGLUT3 Expressing Low-Threshold Mechanosensitive C Fiber Terminals in the Rodent Spinal Cord

doi: 10.1523/ENEURO.0007-19.2019

Figure Lengend Snippet: Primary antibodies used in this study

Article Snippet: VGLUT1 , Mouse, IgG 2b , CL2754 , Human aa 264–293 , Atlas Antibodies , AMAb91041 , AB_2665777 , 1:1000.

Techniques: Concentration Assay

Journal: Ecotoxicology and environmental safety

Article Title: Influence of bifenthrin exposure at different gestational stages on the neural development.

doi: 10.1016/j.ecoenv.2023.115365

Figure Lengend Snippet: Fig. 5. Effect of BF treatment at GD 16-B on the protein levels of VGluT1, VGAT, and NMDA receptors in mouse hippocampi at 6-week-old. (A) Repre sentative western blot images of VGluT1, VGAT, NR1, NR2A and NR2B expression in the mouse hippocampi at 6-week-old. These western blot images are from one membrane. Taking β-Tubulin as loading control. (B) & (C) Quantization of the protein levels of VGluT1, VGAT, VGluT1/VGAT, NR1, NR2A and NR2B of the male mouse (B) and female mouse hippocampi (C) at 6-week-old. N = 4 (litter). The bands in each column of Fig. 5A are from the same pup. Unpaired Student’s t-test was used to analyze the statistical significance. * * P < 0.01 vs. respective sex control.

Article Snippet: Primary antibodies against VGluT1 (#CL2754), VGAT (#131003), NMDA Receptor 2B (NR2B, GluN2B, #21920–1-AP) were bought from Novus Biologicals (Littleton, CO, USA), synaptic systems (Gottingen, Germany), and Proteintech (Chicago, IL, USA) respectively.

Techniques: Western Blot, Expressing, Membrane, Control