Quantifying Mabc-Induced Lung Inflammation

C57BL/6 mice lung tissues were collected after 21 days of Mabc infection. Lung tissues were fixed in 10% formalin, embedded in paraffin wax, and cut into 4 μm sections. The lung sections (4 μm) were stained with hematoxylin and eosin (H&E). Slides were digitalized by scanner PANNORAMIC 300 Flash DX device (3DHISTECH Ltd), and the inflamed area was quantified by using Fiji software. For IHC staining, lung paraffin sections (4 μm) were stained with 4′,6-diamidino-2-phenylindole DAPI (P36935; Invitrogen), anti-NOS2 mouse (sc-7271; Santa Cruz Biotechnology), anti-F4/80 rat (sc-52664; Santa Cruz Biotechnology), and matched secondary antibodies. (A11029 and A21209; Invitrogen) Immunochemical staining images were obtained by a Leica TCS SP8 microscope system (Leica) [29 (link)].

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Jeon S.M., Kim Y.J., Nguyen T.Q., Cui J., Thi Bich Hanh B., Silwal P., Kim J.K., Kim J.M., Oh D.C., Jang J, & Jo E.K. (2022). Ohmyungsamycin promotes M1-like inflammatory responses to enhance host defence against Mycobacteroides abscessus infections. Virulence, 13(1), 1966-1984.

Publication 2022

4′,6-diamidino-2-phenylindole DAPI ( P36935; A11029 A21209 Leica TCS SP8 microscope system

Antibodies C57bl mice Dapi Device Eosin Formalin Infection Lung Microscope Mouse Nos2 Paraffin Tissues

Corresponding Organization : Seoul National University

Other organizations : Keimyung University

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Variable analysis

independent variables

Mabc infection
Duration of Mabc infection (21 days)

dependent variables

Inflamed area in lung tissues
Expression of NOS2
Expression of F4/80

control variables

Mouse strain (C57BL/6)
Lung tissue preparation (fixed in 10% formalin, embedded in paraffin wax, cut into 4 μm sections)
Staining techniques (H&E, DAPI, anti-NOS2, anti-F4/80, and matched secondary antibodies)

controls

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

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