Journal: Cell reports

Article Title: The C9ORF72 repeat expansion alters neurodevelopment

doi: 10.1016/j.celrep.2023.112983

Figure Lengend Snippet: (A) Representative images of brains of 2-month-old mice exposed to vehicle or 2 μg/g VIP antagonist from E9.5–E11.5. (B) Quantification of brain mass in 2-month-old mice that were treated with vehicle or VIP antagonist from E9.5–E11.5. Each data point represents the brain mass of one mouse (n = 9 vehicle and n = 8 VIP antagonist). Mean ± SEM. Unpaired t test. (C and D) Representative images (C) and quantification (D) of cortical thickness in E18.5 control and C9ORF72 -BAC embryos treated with vehicle or 2 μg/g VIP antagonist using immunohistochemistry. Each data point represents the average cortical thickness in one embryo (n = 3 control and 3 C9ORF72 -BAC embryos). The average cortical thickness was determined by measuring the width from layer 1 to layer 6 across 3 different 16-mm sections beginning at approximately bregma + 3.51. Mean ± SEM. One-way ANOVA. Scale bars, 1 mm. (E) Quantification of time to fall in a hanging wire test for 2-month-old control and C9ORF72 -BAC mice treated with vehicle or VIP antagonist from E9.5–E11.5. Each data point represents the average time to fall for one mouse. Control + vehicle, n = 11 mice; control + VIP antagonist, n = 10 mice; C9ORF72 -BAC + vehicle, n = 10 mice; C9ORF72 -BAC + VIP antagonist, n = 16 mice. Mean ± SEM. Kruskal-Wallis test. (F) Quantification of total distance traveled during an open field test for 2-month-old control and C9ORF72 -BAC mice treated with vehicle or VIP antagonist from E9.5–E11.5. Each data point represents the total distance traveled for one mouse. Control + vehicle, n = 11 mice; control + VIP antagonist, n = 10 mice; C9ORF72 -BAC + vehicle, n = 10 mice; C9ORF72 -BAC + VIP antagonist, n = 16 mice. Mean ± SEM. two-way ANOVA.

Article Snippet: Pregnant dams were administered intraperitoneal (IP) injections of 2 μg/g body weight of the VIP receptor antagonist compound (Tocris, cat. no. 3054) once a day at E9.5, E10.5, and E11.5.

Techniques: Control, Immunohistochemistry

Journal: Molecular therapy : the journal of the American Society of Gene Therapy

Article Title: VIP-expressing dendritic cells protect against spontaneous autoimmune peripheral polyneuropathy.

doi: 10.1038/mt.2014.77

Figure Lengend Snippet: Figure 1 Generation and characterization of LV-VIP-DC. (a) Schematic representation of the LV cassette carrying VIP gene (LV-VIP). EF-1α promoter drives expression of hVIP cDNA in pSMPUW-Neo LV plasmid. (b) VIP secretion by LV-VIP–transduced dendritic cells (LV-VIP-DCs). LV-VIP-DCs and controls were cultured at a concentration of 1 × 106 cells/well for 24 hours, and the culture supernatant was analyzed for VIP levels using enzyme-immunoassay method. Error bars represent standard error of the mean (SEM) of four experiments. LV-DC, DCs transduced LV without hVIP cDNA. (c) Flow cytometry showing LV-VIP-DCs–expressing phenotypic markers, CD11c, CD80, CD40, and MHC-II at 3–4-day post- transduction. (d) Quantification of phenotypic markers, LV-VIP-DCs versus controls. Error bars represent SEM of five experiments. Statistical significance: *P < 0.05, **P < 0.01; one-way analysis of variance followed by Tukey’s multiple comparison test. cPPT, central polypurine tract; DC, dendentric cell; EF, elongation factor; hVIP, human vasoactive intestinal polypeptide; LTR, long terminal repeat; LV, lentiviral vector; VIP, vasoactive intestinal polypeptide; WPRE, woodchuck hepatitis post-transcriptional regulatory element; ψ, cis acting elements.

Article Snippet: ViraSafe Lentiviral Expression System was purchased from Cell Biolabs (CA). hVIP cDNA was obtained from Origene (SC319768; OriGene Technologies, Rockville, MD).

Techniques: Expressing, Plasmid Preparation, Cell Culture, Concentration Assay, Enzyme-linked Immunosorbent Assay, Flow Cytometry, Transduction, Comparison