vacht  (Novus Biologicals)

Journal: International Journal of Nanomedicine

Article Title: Bupivacaine Nanoparticles Inhibit Triple-Negative Breast Tumor Growth by Suppressing the Noradrenergic Nerves in Tumor Microenvironment

doi: 10.2147/IJN.S515895

Figure Lengend Snippet: Evaluation of neural type in orthotopic 4T1 tumor. ( A ) Representative immunofluorescence images of tumor tissues for the adrenergic nerve marker Tyrosine Hydroxylase (TH), cholinergic nerve marker vesicular acetylcholine transporter (VAChT) and sensory nerve marker calcitonin gene-related peptide (CGRP). Color coding is indicated at the top. Scale bars, 100 μm ( B ) the quantification of TH, CGRP and VAChT density of representative images. Data show mean ± SD; significant differences were calculated using One-way ANOVA followed by Tukey’s post-hoc test. **: P <0.01, *** P <0.001, ****: P <0.0001.

Article Snippet: Antigen retrieval was performed with citrate antigen retrieval solution (Beyotime, P0081) by heating slices in boiling temperature for 5 min. After cooling to room temperature, the slices were were blocked and permeabilized with PBS containing 10% goat serum and 0.3% Triton X-100 for 1 h. Then the slices were incubated with the following primary antibodies at 4°C overnight: F4/80 antibody (1:200, Abcam, ab6640); anti-TH antibody (1:600, Merck, AB152); anti-VAChT antibody (1:200, Novus Biologicals, NB110-74,764); anti-CGRP (1:200, Thermo Fisher Scientific, PA5-114929).

Techniques: Immunofluorescence, Marker

Journal: Biochemical pharmacology

Article Title: CDP-choline modulates cholinergic signaling and gut microbiota to alleviate DSS-induced inflammatory bowel disease.

doi: 10.1016/j.bcp.2023.115845

Figure Lengend Snippet: Fig. 3. Effect of CDP-choline on ACh synthesis-related proteins in colonic tissue of mice. (A) The representative image of ChT1, ChAT, VAChT, AChE, TNF-α, and IL-6 expression among the three groups. Histogram of the relative expressions of ChT1 (B), ChAT (C), VAChT (D), AChE (E), TNF-α (F), and IL-6 (G), respectively (n = 6). (H) Histogram of ChAT activity in the colon among the three groups measured by ELISA (n = 8). Data are expressed as mean ± SEM. * P < 0.05, ** P < 0.01, *** P < 0.001.

Article Snippet: The membranes were sealed (20 °C, 12 min) in protein-free rapid blocking buffer (EpiZyme, PS108P, China) and incubated overnight at 4 °C in the following primary antibody: β-Actin (1:1000, Cell Signaling Technology, #3700, USA), Occludin (1:1000, Cell Signaling Technology, #91131, USA), High-affinity choline transporter (SLC5A7/ChT1 1:1000, ThermoFisher, PA5-42485, USA), Choline acetyltransferase (ChAT, 1:1000, Cell Signaling Technology, #27269, USA), Acetylcholinesterase (AChE, 1:1500, Abcam, ab183591, UK), Vesicular acetylcholine transporter (SLC18A3/VAChT, 1:1000, Boster Bio, A04891, China), α7 nicotinic acetylcholine receptor (CHRNA7/α7 nAChR, 1:1000, Proteintech, 21379–1-AP, China), Tumor necrosis factor-α (TNF-α, 1:1000, Cell Signaling Technology, #11948, USA), Interleukin 6 (IL-6, 1:1000, HUABIO, R1412-2, China) and ZO-1 (1:1500, Bioswamp, PAB36669, China).

Techniques: Expressing, Activity Assay, Enzyme-linked Immunosorbent Assay