Quantifying 53BP1 in Liver Sections

Liver sections from CTRL and IUGR males at birth and 6-month-old rats were stained with 53BP-1 (1/100, Abcam, Cambridge, UK) overnight at 4 °C. Sections were then washed with PBS and incubated for two hours with Alexa Fluor-647-conjugated goat anti-rabbit IgG (1/200, Abcam). Sections were then rinsed with PBS and mounted using Fluoromount g mounting medium with DAPI. A negative control was established through incubating only with secondary antibody. Slides were observed blindly using a fluorescence microscope (Nikon, Eclipse Ti2 Series) by the same experimenter (C.Y.) [30 (link)]. Fluorescence was evaluated with ImageJ software, and liver autofluorescence was subtracted

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Keshavjee B., Lambelet V., Coppola H., Viertl D., Prior J.O., Kappeler L., Armengaud J.B., Chouraqui J.P., Chehade H., Vanderriele P.E., Allouche M., Balsiger A., Sarre A., Peyter A.C., Simeoni U, & Yzydorczyk C. (2022). Stress-Induced Premature Senescence Related to Oxidative Stress in the Developmental Programming of Nonalcoholic Fatty Liver Disease in a Rat Model of Intrauterine Growth Restriction. Antioxidants, 11(9), 1695.

Publication 2022

Alexa Fluor-647-conjugated goat anti-rabbit IgG Eclipse Ti2 Series

Alexa fluor 647 Anti igg Antibody Birth males Dapi Fluorescence Fluorescence microscope Goat Iugr Liver Rabbit Rats

Corresponding Organization : University of Lausanne

Other organizations : Centre de Recherche Saint-Antoine, Fondation pour l’innovation en Cadiométabolisme et Nutrition, Inserm, Sorbonne Université

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Variable analysis

independent variables

Treatment (CTRL vs. IUGR)

dependent variables

53BP-1 fluorescence intensity in liver sections

control variables

Age (birth and 6-month-old rats)
Sex (males)
Liver tissue sections
Staining protocol (53BP-1 primary antibody, Alexa Fluor-647-conjugated secondary antibody, DAPI)
Microscopy (fluorescence microscope, ImageJ software for analysis)

controls

Negative control: Incubation with only secondary antibody

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