Vitamin A Regulation of RAR-Mediated Transcription

Mice were maintained at IMM or VU University Medical Centre according to national and international guidelines. Bone marrow cells were isolated from 8 week-old C57Bl/6 CD45.1 mice and injected intravenously into 2 week-old lethally irradiated CD45.2 ROSA26-RARa403^Het (WT→Rar^Het) or WT littermate controls (WT→WT). C57Bl/6 female mice received either vitamin A deficient (VAD), with no vitamin A (vitamin free casein), vitamin A high (VAH, 25,000IU/Kg) or vitamin A control (VAC, 4,000IU/kg) diets. Retinoic acid was provided to pregnant mice from E10.5 until sacrifice. Quantitative real time RT-PCR was performed as previously described^{6 (link),7 (link),10 (link)}. Computational analysis was performed with TESS. DNA/protein complexes were immunoprecipitated using antibodies against mouse pan-RAR, pan-RXR or control IgG.

Partial Protocol Preview
This section provides a glimpse into the protocol.
The remaining content is hidden due to licensing restrictions, but the full text is available at the following link: Access Free Full Text.

van de Pavert S.A., Ferreira M., Domingues R.G., Ribeiro H., Molenaar R., Moreira-Santos L., Almeida F.F., Ibiza S., Barbosa I., Goverse G., Labão-Almeida C., Godinho-Silva C., Konijn T., Schooneman D., O’Toole T., Mizee M.R., Habani Y., Haak E., Santori F.R., Littman D.R., Schulte-Merker S., Dzierzak E., Simas J.P., Mebius R.E, & Veiga-Fernandes H. (2014). Maternal retinoids control type 3 innate lymphoid cells and set the offspring immunity. Nature, 508(7494), 123-127.

Publication 2014

Antibodies Bone marrow cells C57bl mice Casein Diets Female Mouse Protein Quantitative real time pcr Retinoic acid Tess Vitamin Vitamin a

Corresponding Organization :

Other organizations : Royal Netherlands Academy of Arts and Sciences, Hospital de Egas Moniz, Amsterdam UMC Location VUmc, Erasmus University Rotterdam, New York University, University Medical Center Utrecht

Top 5 similar protocols

Protocol cited in 8 other protocols

Variable analysis

independent variables

Vitamin A deficient (VAD) diet
Vitamin A high (VAH, 25,000IU/Kg) diet
Vitamin A control (VAC, 4,000IU/kg) diet
Retinoic acid provided to pregnant mice from E10.5 until sacrifice

dependent variables

Quantitative real time RT-PCR analysis
DNA/protein complexes immunoprecipitated using antibodies against mouse pan-RAR and pan-RXR

control variables

2 week-old lethally irradiated CD45.2 ROSA26-RARa403Het (WT→RarHet) or WT littermate controls (WT→WT)
Mice maintained at IMM or VU University Medical Centre according to national and international guidelines
Bone marrow cells isolated from 8 week-old C57Bl/6 CD45.1 mice

positive controls

WT littermate controls (WT→WT)

negative controls

Control IgG for DNA/protein complex immunoprecipitation

Annotations

Based on most similar protocols

Etiam vel ipsum. Morbi facilisis vestibulum nisl. Praesent cursus laoreet felis. Integer adipiscing pretium orci. Nulla facilisi. Quisque posuere bibendum purus. Nulla quam mauris, cursus eget, convallis ac, molestie non, enim. Aliquam congue. Quisque sagittis nonummy sapien. Proin molestie sem vitae urna. Maecenas lorem.

As authors may omit details in methods from publication, our AI will look for missing critical information across the 5 most similar protocols.

About PubCompare

Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.

We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.

However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.

Ready to get started?

Revolutionizing how scientists
search and build protocols!