Histological Analysis of Mouse Middle Ear

For histological analysis, formalin-fixed paraffin-embedded mouse middle ear tissues were sectioned (4–5 μm), and then stained with hematoxylin and eosin (H&E) to visualize inflammatory responses and pathological changes in the middle ear as described previously (32 (link), 33 (link), 35 (link)). For IF assay, IF detection of CYLD protein was performed using mouse anti-CYLD (Santa Cruz Biotechnology), and FITC-conjugated goat anti-mouse IgG (Santa Cruz Biotechnology) in the paraffin section of mouse middle ear tissues as described previously (35 (link)). Images of stained tissue sections were recorded under light- and fluorescence-microscopy systems (AxioVert 40 CFL, AxioCam MRC, and AxioVision LE Image system, Carl Zeiss), and analyzed by using a quantitative image analysis system with the VISIOPHARM Integrator System software version 3.0.8.0 (Visiopharm, Horsholm, Denmark).

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Komatsu K., Nam D.H., Lee J.Y., Yoneda G., Yan C, & Li J.D. (2020). Vinpocetine suppresses Streptococcus pneumoniae-induced inflammation via inhibition of ERK1 by CYLD. Journal of immunology (Baltimore, Md. : 1950), 204(4), 933-942.

Publication 2019

FITC-conjugated goat anti-mouse IgG AxioVert 40 CFL AxioCam MRC AxioVision LE Image system VISIOPHARM Integrator System software

Anti igg Assay Cyld Cyld protein Eosin Fitc Fluorescence microscopy Formalin Goat Inflammatory Light Middle ear Mouse Paraffin Stained tissue Tissues

Corresponding Organization : Georgia State University

Other organizations : Chung-Ang University, University of Rochester Medical Center

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Variable analysis

independent variables

Mouse middle ear tissues

dependent variables

Inflammatory responses and pathological changes in the middle ear
CYLD protein expression

control variables

Formalin-fixed paraffin-embedded tissue sections
Hematoxylin and eosin (H&E) staining
Immunofluorescence (IF) detection of CYLD protein using mouse anti-CYLD and FITC-conjugated goat anti-mouse IgG
Light- and fluorescence-microscopy imaging systems
Quantitative image analysis using VISIOPHARM Integrator System software

controls

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

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