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, Supplemental Information The SCRIB Paralog LANO/LRRC1 Regulates Breast Cancer Stem Cell Fate through WNT/b-Catenin Signaling

L. Lopez-almeida, M. Sebbagh, F. Bertucci, P. Finetti, J. Wicinski et al., Santoni The (Lano -/-) were characterized by PCR and sequencing of the remaining genomic region. Finally, absence of Lano protein expression was tested biochemically on recombinant tissues. Primers used for routine genotyping of mice biopsies are the following: Cre detection


. Lanowt,


. Lanocko,

, Lano/LRRC1 excised gene [LanoCKO(1F)

. Lanowt,

, Mice were housed under sterile conditions with sterilized food and water provided ad libitum and maintained on a 12-h light and 12-h dark cycle. 1×10 6 luciferase-expressing SUM149 cells suspended in 50% phenol red-free Matrigel (Becton Dickinson Bioscience) were inoculated in the mammary fat pad in 3 cohorts of 8 mice. Tumor growth was monitored by measuring with a digital caliper and calculating tumor volume (length × width 2 × ?/6). All animals were randomly assigned to treatment groups, All experiments were performed in agreement with the French Guidelines for animal handling and approved by local ethics committee APAFIS#10719-2017071709337799. NOD/SCID/?c null mice (NSG) were obtained from Charles River

. Photonimager, After completion of the analysis, autopsy of mice was performed, and organ luminescence was assessed. For the limiting dilution assay, 100; 1,000; 100,000 cells were injected as described above in the mammary fat pad of NGS mice corresponding (9 cohorts of 4 mice). Estimation of the breast CSC frequency was calculated using ELDA algorithm

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