Immunofluorescence analysis of brain slices

After 7‐day co‐treatment of LPS and natural compounds, the mice were perfused with saline and 4% paraformaldehyde for IF. A total of 30 μm‐thick coronal brain slices (AP −2.0 and −2.5 mm from bregma) were obtained using cryomicrotome (Thermo Fisher). IF was conducted as previously reported,
⁶ (link) with the following primary antibodies: anti‐GFAP (chicken polyclonal, 1:1000; MilliporeSigma, AB5541), anti‐GABA (guinea pig polyclonal, 1:500; MilliporeSigma, AB175) and anti‐MAO‐B (rabbit polyclonal, 1:1000; Abcam, AB137778). Confocal images (60x) were obtained with a multiphoton laser scanning microscope (Nikon A1R, Tokyo, Japan) and analyzed by Imaris 9.3 software.

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Cho J., Hong E., Kim Y., Song J., Ju Y.H., Kim H., Lee H., Kim H, & Nam M. (2024). Baicalin and baicalein from Scutellaria baicalensis Georgi alleviate aberrant neuronal suppression mediated by GABA from reactive astrocytes. CNS Neuroscience & Therapeutics, 30(5), e14740.

Publication 2024

cryomicrotome AB5541 AB175

Corresponding Organization :

Other organizations : Korea Institute of Brain Science, Korea Institute of Science and Technology, Woosuk University, Kyung Hee University, Korea University of Science and Technology, Bio-Medical Science (South Korea)

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Variable analysis

independent variables

LPS (lipopolysaccharide)
Natural compounds

dependent variables

GFAP (glial fibrillary acidic protein) expression
GABA (gamma-aminobutyric acid) expression
MAO-B (monoamine oxidase B) expression

control variables

Brain region (coronal brain slices from AP -2.0 and -2.5 mm from bregma)
Slice thickness (30 μm)
Perfusion method (saline and 4% paraformaldehyde)
Imaging technique (confocal microscopy, 60x)
Image analysis software (Imaris 9.3)

controls

Positive control: Not explicitly mentioned
Negative control: Not explicitly mentioned

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