Analyzing Notch Signaling in Mice

The Columbia University Institutional Animal Care and Use Committee approved protocols used in animal studies. All mice were maintained on a C57BL/6 background. For assessment of wild type expression patterns, we used C57BL/6J virgin female mice and males of proven fertility (The Jackson Laboratory). The CBF:H2B-Venus transgenic mouse strain that expresses human histone H2B fused to yellow fluorescent protein (YFP) Venus in response to Notch/CSL transcriptional activation was used to determine Notch activity [38 (link)]. Mice were bred; noon on the day a mating plug was observed was designated embryonic day (E) 0.5. Pieces of uteri and implantation sites from pregnant females at E3.5 and E6.5, respectively, were embedded in Tissue-Tek® O.C.T.™ Compound (Sakura Fine Technical Co, Ltd, Tokyo, Japan), snap-frozen on dry ice in ethanol and stored at −80 °C in methylbutane (Sigma-Aldrich).

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Shawber C.J., Lin L., Gnarra M., Sauer M.V., Papaioannou V.E., Kitajewski J.K, & Douglas N.C. (2015). Vascular Notch proteins and Notch signaling in the peri-implantation mouse uterus. Vascular Cell, 7, 9.

Publication 2015

methylbutane

Animal C57bl mice Care protocols Dry ice Embryonic Ethanol Female Fertility Frozen Histone h2b Human Implantation Males Methylbutane Mice Pregnant females Protein Strain Tissue Transcriptional activation Transgenic mouse Uteri

Corresponding Organization :

Other organizations : Columbia University Irving Medical Center

Top 5 similar protocols

Variable analysis

independent variables

Notch/CSL transcriptional activation

dependent variables

Expression patterns of wild type mice
Notch activity

control variables

C57BL/6J virgin female mice
C57BL/6 male mice of proven fertility

controls

CBF:H2B-Venus transgenic mouse strain expressing human histone H2B fused to yellow fluorescent protein (YFP) Venus as a positive control for Notch activity

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