Journal: International Journal of Medical Sciences

Article Title: Parvovirus B19 Nonstructural Protein-Induced Damage of Cellular DNA and Resultant Apoptosis

doi:

Figure Lengend Snippet: PARP is active and necessary for efficient apoptosis in NS1-transfected cells. A. Immunoprecipitated GFP/NS1 protein was poly(ADP ribose)ylated, as shown by a band at 100 kd on the blot probed with anti-poly ADP ribose (PAR, left blot) antibodies. The blots were stripped and reprobed with anti-GFP (right), showing that PAR colocalizes with the GFP/NS1 fusion protein. GFP alone is not ribosylated, as evidenced by the lack of a PAR band corresponding to GFP at 37 kD. Blots shown are representative of three independent experiments. B. PARP activity is necessary for optimal NS1-induced apoptosis. Addition of the PARP inhibitor 5-aminoisoquinolinone (5AIQ) to GFP/NS1 expressing HepG2 cells reduced apoptosis by 57% ( p <0.003). Addition of 5-aminoisoquinolinone had no effect on the GFP transfected cells. N=3, error bars indicate the range of values.

Article Snippet: 25 μl of anti-GFP polyclonal antibody (Rockland, Gilbertsville, PA) were added and the mixture was allowed to bind for 14 hours at 4 o C in an end-over end rotator.

Techniques: Transfection, Immunoprecipitation, Activity Assay, Expressing

Journal: Heliyon

Article Title: Median raphe glutamatergic neuron-mediated enhancement of GABAergic transmission and suppression of long-term potentiation in the hippocampus

doi: 10.1016/j.heliyon.2024.e38192

Figure Lengend Snippet: MRN VGLUT3 neuron-mediated glutamatergic transmission in SR/SLM 5-HT3aR neurons. A) A schematic of AAV injection into the MRN. The left panel shows an image of the MRN from VGLUT3 Cre/5-HT3aR-EGFP mouse which received an AAV-EF1a-double floxed-hChR2(H134R)-mCherry-WPRE-HGHpA injection into the MRN. Scale 150 μm. B) An image of the hippocampus from MRN VGLUT3 ChR2/5-HT3aR-EGFP mouse. The inset shows EGFP-expressing SR/SLM 5-HT3aR neurons and mCherry-expressing MRN VGLUT3 axons. Scale 300 μm/25 μm (inset). C) The upper panel shows a schematic for recording MRN VGLUT3 neuron-mediated transmission in SR/SLM 5-HT3aR neurons. The bottom panel shows example traces for MRN VGLUT3 neuron-mediated EPSCs in SR/SLM 5-HT3aR neurons without drug application, in the presence of tetrodotoxin (TTX, 1 μM), TTX+4-aminopyridine (4-AP, 100 μM) and TTX+4-AP + DNQX (10 μM). Scale 50 pA/10 ms. The blue line indicates light application. D) EPSC amplitude in SR/SLM 5-HT3aR neurons before the drug application, in the presence of TTX, TTX+4-AP, and TTX+4-AP + DNQX (6 neurons from 3 female mice and 6 neurons from 3 male mice). F (1,11) = 22.01, P = 0.001. Data are presented as mean ± SEM. E) EPSC amplitude in female (17 neurons/5 mice) and male (16 neurons/5 mice) groups. Female and male groups did not show statistically significant difference in EPSC amplitude (P = 0.8). The horizontal line in each group represents the mean and the vertical line represents SEM. F) The delay between the onset of the light stimulation and the onset of EPSCs in female (17 neurons/5 mice) and male (16 neurons/5 mice) groups. Female and male groups did not show a statistically significant difference in the delay between the onset of light stimulation and the onset of EPSCs (P = 0.77). n.s. not statistically significant.

Article Snippet: The slices were then incubated in mCherry polyclonal antibody (Rockland Immunochemicals, Cat. No. 600401P16) or Anti-GFP antibody (Abcam, Cat. No. ab13970, RRID: AB_300798 ) at a dilution of 1:1000 in 0.3 % Triton-X/TBS for 24 h at 4 °C [ , ].

Techniques: Transmission Assay, Injection, Expressing

Journal: Journal of Cellular and Molecular Medicine

Article Title: Bioengineered AAV9 and Optimised Microdystrophin Vectors Augment Phenotypic Rescue in a Murine Model of Duchenne Muscular Dystrophy

doi: 10.1111/jcmm.71078

Figure Lengend Snippet: Validation of codon‐optimised microdystrophin transgene in vitro. Transduction of AAV9K51Q‐μDys and AAV9K51Q‐CoμDys vectors in C2C12 cells (A), at an MOI: 1 × 10 5 and its quantification (B) are shown. Representative images were obtained in confocal microscope (AXR Nikon) with a 40X objective (Scale bar: 50 μm). Semi‐quantification was performed from images ( n ≥ 8 per group) obtained in ZOE‐Fluorescent cell imager. The data is represented as mean ± SD. Asterisk (*) represents statistical comparison of AAV9K51Q‐μDys and AAV9K51Q‐CoμDys treated conditions with respect to the cell control and Hash (#) refers to statistical comparison of AAV9K51Q‐CoμDys with respect to AAV9K51Q‐μDys. # p < 0.05, *** p < 0.001, **** p < 0.0001. For statistical comparison, a one‐way ANOVA was performed (GraphPad Prism 8.0.2 software).

Article Snippet: The images were obtained in the ZOE‐Fluorescent cell imager (Bio‐Rad, Hercules, CA, USA) and these images ( n ≥ 8/group) were further used for semi‐quantification analysis (ImageJ software).

Techniques: Biomarker Discovery, In Vitro, Transduction, Microscopy, Comparison, Control, Software