Evaluating Liver Tissue Pathways via Immunohistochemistry

The statuses of lipid peroxidation, M1/M2 macrophage polarization, necroptosis, and pyroptosis were determined through immunohistochemistry staining, per a previously reported method [7 ,8 (link),9 (link),10 (link),11 (link)]. Liver tissues (in paraffin sections) were incubated with primary antibodies against one of the following: malondialdehyde (MDA; Abcam), a lipid peroxidation-related protein [33 (link)]; inducible nitric oxide synthase (iNOS; Abcam), an M1 phase macrophage polarization-related protein [34 (link)]; CD206 (Abcam), an M2 phase macrophage polarization-related protein [34 (link)]; phosphorylated mixed lineage kinase domain-like pseudokinase (pMLKL; Abcam), a necroptosis-related protein [35 (link)]; or NLRP3 (Abcam), a pyroptosis-related protein [17 (link)]. All the sections were observed under the TissueGnostics Axio Observer Z1 microscope (TissueGnostics, Vienna, Austria) and then quantified and analyzed using the image processing software Image J.

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Shih H.J., Chang C.Y., Chiang M., Le V.L., Hsu H.J, & Huang C.J. (2021). Simultaneous Inhibition of Three Major Cytokines and Its Therapeutic Effects: A Peptide-Based Novel Therapy against Endotoxemia in Mice. Journal of Personalized Medicine, 11(5), 436.

Publication 2021

CD206 NLRP3 Axio Observer Z1 microscope

Antibodies Inducible nitric oxide synthase Inos Lipid peroxidation Liver Macrophage polarization Malondialdehyde Microscope Mixed lineage kinase Necroptosis Paraffin Protein Pyroptosis Tissues

Corresponding Organization : Tzu Chi University

Other organizations : Changhua Christian Hospital, Hue University

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Protocol cited in 2 other protocols

Variable analysis

independent variables

Immunohistochemistry staining protocol

dependent variables

Statuses of lipid peroxidation
M1/M2 macrophage polarization
Necroptosis
Pyroptosis

control variables

Liver tissues (in paraffin sections)

controls

Positive control: None specified
Negative control: None specified

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