Evi1 mRNA in situ hybridization was carried out using a full length Evi1 cDNA probe [35] (link) using standard protocols. Probes were labeled using a DIG RNA Labeling Kit (Roche Applied Science, Tokyo, Japan). Detection was via an anti-DIG antibody coupled to alkaline phosphatase (Roche, Tokyo, Japan) followed by staining with BCIP-NBT (Bromo-4-chloro-3-indolyl Phosphate/Nitro Blue Tetrazolium) (Nacalai, Tokyo, Japan) as previously described [36] (link).
Bard-Chapeau E.A., Szumska D., Jacob B., Chua B.Q., Chatterjee G.C., Zhang Y., Ward J.M., Urun F., Kinameri E., Vincent S.D., Ahmed S., Bhattacharya S., Osato M., Perkins A.S., Moore A.W., Jenkins N.A, & Copeland N.G. (2014). Mice Carrying a Hypomorphic Evi1 Allele Are Embryonic Viable but Exhibit Severe Congenital Heart Defects. PLoS ONE, 9(2), e89397.
Other organizations :
Institute of Molecular and Cell Biology, Wellcome Centre for Human Genetics, National University Cancer Institute, Singapore, Yale University, University of Rochester Medical Center, RIKEN Center for Brain Science, Inserm, Université de Strasbourg, Centre National de la Recherche Scientifique
DIG RNA Labeling Kit (Roche Applied Science, Tokyo, Japan)
Anti-DIG antibody coupled to alkaline phosphatase (Roche, Tokyo, Japan)
BCIP-NBT (Bromo-4-chloro-3-indolyl Phosphate/Nitro Blue Tetrazolium) (Nacalai, Tokyo, Japan)
controls
Positive control: Not explicitly mentioned
Negative control: Not explicitly mentioned
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